AQUACULTURE HEALTH I N T E R N A T I O N A L

ISSUE 7 NOVEMBER 2006 NZ$10.00

STREPTOCOCCUS
IN WARM-WATER FISH

PROBIOTIC BACTERIA’S
EFFECT ON SHRIMP SURVIVAL

Aquaculture without FEATURED DIAGNOSTIC

Frontiers seeks help LABORATORY

CANNIBALISM
in juvenile Atlantic cod
CONTENTS ISSUE 7, NOVEMBER 2006

3 EDITORIAL
Are mergers and acquisitions good for fish health?
4 FACILITY FOCUS
Research and development at Clear Springs Foods
7 FOCUS ON FINFISH
Bacterial haemorrhagic septicaemia in tilapias
8 BOOK REVIEW
Atlas of Tilapia Histology 2006
10 10 AN OVERVIEW
Streptococcus in warm-water fish
16 NEWS
Updates from around the globe
22 FACILITY FOCUS
Fish health investigations at Ardtoe Marine Laboratory, Scotland
24 DIAGNOSTIC LABORATORY SERIES
Veterinary Diagnostic Service, Institute of Aquaculture,
Stirling University

19 28 RESEARCH
Probiotic bacteria’s effect on shrimp survival
30 PERSPECTIVES
Coarse fisheries and fish health in England
32 BOOK REVIEW
Systemic Pathology of Fish AQUACULTURE
HEALTH I N T E R
N A T I O
ISSUE 7 NOVE N A L
MBER 2006
NZ$10 .00

STREPTOCO
IN WARM-WA CCUS
TER FISH
PROBIOTIC BAC
EFFECT ON SHR TERIA’S
Aquaculture with IMP SURVIVAL
Frontiers seeks out FEATURED DIA
help GNOSTIC
LABORATORY

ON THE
24 COVER
Fish Health
Investigations at
Ardtoe Marine
Laboratory, Scotland
See page 22.
CANNIBALISM
in juvenile A
tlantic cod

EDITORIAL DIRECTOR: Dr Scott Peddie CONTRIBUTORS:

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2 AQUACULTURE HEALTH INTERNATIONAL NOVEMBER 2006

 EDITORIAL
ARE MERGERS & ACQUISITIONS
GOOD FOR FISH HEALTH?
DR SCOTT PEDDIE, EDITORIAL DIRECTOR

T
he news this week emanating from the United Kingdom’s
competition authority, the Competition Commission, is
that provisional clearance has been granted for Pan Fish’s
proposed acquisition of Marine Harvest.
In a summary of provisional findings released on November
7, the commission indicated that it had reached the conclusion
that the acquisition would not result in a substantial lessening of
competition.
Pan Fish’s acquisition of Marine Harvest, together with Fjord
Seafood, now makes it the largest salmon farming company in the
world by far, accounting for approximately 25 percent of global
harvest volume (using 2006 figures).
Of course acquisitions and mergers are nothing new to a
global salmon farming industry where consolidation is a by-word
for survivability in an increasingly competitive marketplace.
Economies of scale make larger and larger farming units, and
hence larger and larger companies, inevitable. Such consolidation
ensures that the industry remains viable and continues to employ
large numbers of people in economically fragile and often
geographically isolated areas.
But what about fish health? Recent changes in the salmon
farming industry have led me to ponder on the knock-on
effects of consolidation on this important aspect of production
management. Thankfully, it seems to me that in the fish welfare
and health arena, the effects are overwhelmingly positive.
Larger companies typically invest
more money in fish health programmes
and expertise and related research and
development. They also tend to lead
the way in developing best practices
in fish health management at a very
practical level.
Not only that, it would seem logical to
assume that the increasing prominence
of Area Management Agreements and the like will be aided by a
reduction in the number of companies operating in particular
localities. More consistent strategic management in this respect
can only be a good thing.
I wonder also how the pharmaceutical companies will view
an increasingly consolidated global industry? Competition in
this market is very tight indeed, and is therefore dominated
by a small number of key players. One can only assume
that competition will increase in the relatively small market
segments where companies have products that compete directly
for market share.
So, as always, changes in industry structures result in numerous
opportunities for positive developments, and in this respect the
fish health arena is no different. One thing’s for sure - industry
consolidation should make for interesting developments in the
months and years ahead! ■

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NOVEMBER 2006 AQUACULTURE HEALTH INTERNATIONAL 3

FAC I L I T Y F O C U S
RESEARCH AND DEVELOPMENT
AT CLEAR SPRINGS FOODS
BY DR SCOTT E LAPATRA
RESEARCH DIVISION, CLEAR SPRINGS FOODS, INC, BUHL, IDAHO, UNITED STATES
C
lear Springs Foods Inc is an employee-owned, vertically
integrated company producing high quality rainbow trout
for human consumption. Clear Springs Foods is composed
of several divisions, including operations (broodstock, feed mill,
fish production, processing and distribution), marketing, sales,
administration, and research and development.
The research and development centre is located next to our
research hatchery and the Jess O “Ted” Eastman Visitor Centre in the
Snake River Canyon in southern Idaho, USA.
The research and development division conducts primarily applied
research focused on optimising fish production and ensuring
WATER QUALITY ANALYSES ARE
CONDUCTED ACCORDING TO
GOOD LABORATORY PROCEDURES,
INCLUDING A QUALITY ASSURANCE
PROGRAMME
4 AQUACULTURE HEALTH INTERNATIONAL NOVEMBER 2006
THE RESEARCH AND DEVELOPMENT CENTRE IS
NEXT TO OUR RESEARCH HATCHERY AND THE JESS
O “TED” EASTMAN VISITOR CENTRE IN THE SNAKE
RIVER CANYON IN SOUTHERN IDAHO
consistent, high quality trout are produced economically and in an
environmentally responsible manner.
Work areas include environmental science, fish health management,
fish culture, fish production and breeding, and fish nutrition.
As a resource-based industry we are committed to responsible
environmental stewardship.
Our rainbow trout are produced in the high quality water
of the Snake Plain aquifer. This water quality and volume is
routinely monitored. Effluent water quality is continually being
characterised, and techniques to further minimise fish production
impact on the environment developed. Two “dry” water quality
laboratories are devoted to this effort. All analyses are conducted
according to good laboratory procedures, including a quality
assurance programme.
Maintenance of fish health is essential for optimal rainbow trout
production. We do this by identifying potential problems and
developing strategies to minimise any impact they may have. These
strategies range from formulating effective vaccines, to improvements
in fish culture techniques such as biosecurity.
The basic strategy is to first diagnose a problem, then develop
a solution and test the solution under controlled environmental
conditions. After successful laboratory evaluation, solutions are
field tested and fine-tuned. We are constantly striving to refine our
production programme through research on photo-period control,
nutrition and selective breeding.
SELECTIVE BREEDING
Selective breeding of rainbow trout is an important component of
our r and d programme. The current goals of the selective breeding
programme are to improve growth and disease resistance. To
improve these traits, data is recorded on thousands of individuals
each year. Growth data is collected at various ages to determine

THE SPECIFIC-
PATHOGEN-FREE
LABORATORY, WHERE
SELECTED RAINBOW
TROUT FAMILIES ARE
INCUBATED AND HATCHED
which families and which individuals within each family have
the best growth. To improve disease resistance, a portion of the
progeny from each family is exposed to specific pathogens in
a standardised challenge test. Currently, each family is tested
for survivability to infectious hematopoietic necrosis virus and
Flavobacterium psychrophilum, the causative agent of bacterial
coldwater disease and rainbow trout fry syndrome.
Selection to improve growth began when the breeding programme
was initiated. The average weight of the odd-year generation group
increased from 660g at 328 days of age in 1991 to 921g at 301 days in
2003. The average weight of the even-year group increased from 620g
at 328 days in 1992 to 866g at 301 days in 2004.
Selection to improve IHN resistance started with the 1994
generation. Using a standardised challenge test, IHN mortality
decreased by 25.8 percent in the odd-year generation group and 29.7
percent in the even-year group. Growth is a moderately heritable
trait that can be changed rapidly and economically with traditional
quantitative genetic techniques. (See photo above.)
GENE EXPRESSION
Studies involving fish growth and disease resistance rely heavily
on monitoring physical parameters such as overall weight gain
and fillet weight, or mortality after exposure and immunological
activity. These measurements do provide valuable information,
in a general manner, on variation between control and treatment
groups.
The recent substantial increase in genome sequence information
for trout and other species now allows for the generation of assays
for the analysis of expression for genes of interest. These assays can
be used to study the effects of immunological stimulus on genes
of known function, or to develop assays that correlate expression
changes with known changes in animal physiology.
In collaboration with Dr Ken Overturf (United States Department
of Agriculture-Agricultural Research Service, Hagerman, Idaho),
we have developed a number of real-time quantitative PCR assays
for examining the expression of genes involved with muscle
development, immunology and metabolism.
In some experiments there is a high correlation between the
level of gene expression and treatment. We are now using these
assays to study how immune stimulation affects muscle growth in
aquaculture species. Specifically, we are examining what immune
genes are stimulated by specific pathogens and how these, and
immuno-stimulants added into diets, affect muscle growth.
THE SPECIFIC-PATHOGEN-INFECTED LABORATORY, WHERE
SELECTED FAMILIES ARE EVALUATED FOR SURVIVABILITY TO CERTAIN
PATHOGENS, AND POTENTIAL VACCINES ARE EVALUATED
DNA VACCINATION
Dr Gael Kurath’s research group (United States Geological Survey,
Seattle, Washington) has developed a highly efficacious experimental
DNA vaccine against IHNV, based on the viral glycoprotein (G) gene,
and we have collaborated extensively on characterising this vaccine.
(See photo above.)
Rainbow trout fry injected intra-muscularly (i.m.) with small
quantities of vaccine plasmid DNA (0.1µg) are protected against
virulent IHNV challenge for up to two years post-vaccination. The
vaccine induces three distinct phases of response: an early anti-viral
response (EAVR), a specific anti-viral response (SAVR) and a long-
term anti-viral response (LAVR).
The three phases differ in the specificity of protection, the
magnitude of relative percent survival of vaccinated animals in a
virulent IHNV challenge, and neutralising antibody prevalence and
titer.
Dr Jim Winton’s research group (USGS, Seattle, Washington) used
a combination of micro-array and quantitative reverse transcriptase
PCR (qRT-PCR) methodology to define the host responses during
the EAVR phase.
Increased expression of a large number of immune-related genes
was observed at the primary site of vaccine delivery, while only
the type I interferon-related genes were up-regulated in secondary
systemic sites. These studies emphasise the potent nature of the
IHNV viral glycoprotein to stimulate innate anti-viral immunity.
Many questions still remain regarding immune mechanism(s)
involved in the EAVR, SAVR and LAVR, and research strategies are
being developed to address these questions. One of these studies
examined whether administering exogenous antigen in the form of
a whole, killed IHN virus vaccine could elicit an EAVR in rainbow
trout similar to that elicited by the DNA vaccine.
The killed IHN vaccines were prepared by Drs Eric Anderson
and Sharon Clouthier (Maine BioTek Inc, Winterport, Maine)
by the inactivation of IHN virus with formaldehyde, binary
ethylenimine (BEI) or propiolactone. We immunised rainbow trout
by intraperitoneal injection with propiolactone inactivated IHN
virus, and were able to elicit a potent EAVR 105 degree days after
vaccination, with relative percent survival (RPS) values of between
80 and 100 percent.
The formaldehyde and BEI inactivated IHN virus vaccines also
elicited an early antiviral response in immunised rainbow trout,
though the RPS values were 50 to 80 percent. These results indicate
that vaccination of rainbow trout with exogenous IHN virus
NOVEMBER 2006 AQUACULTURE HEALTH INTERNATIONAL 5
FAC I L I T Y F O C U S
antigen(s) can elicit an EAVR comparable to that elicited by the DNA
vaccine, and that the inactivating agent used to prepare the antigen
can alter the efficacy of the response.
As DNA vaccination is a relatively new technology, various
theoretical and long-term safety issues related to the environment
and the consumer remain to be fully addressed, although inherently
the risks should not be any greater than with the commercial fish
vaccines that are currently used.
Present classification systems lack clarity in distinguishing DNA-
vaccinated animals from genetically modified organisms, which
could raise issues in terms of licensing and public acceptance of the
technology. A study recently examined the effect on vaccine efficacy
of using alternative i.m. injection sites that would ultimately be
removed from the fish during processing before the product goes to
market. The results indicate that alternative sites are available that do
not effect the efficacy of the vaccine.
FISH PERFORMANCE
It has also been hypothesised that vaccination with a potent biologic
could negatively impact fish growth and performance. The IHNV
DNA vaccine was used to test this hypothesis.
Triplicate 30-fish groups of rainbow trout with a mean weight
of 50g were either injected i.m. with 10µg of the DNA vaccine, or
mock-vaccinated and fed approximately 1.8 percent body weight per
day ration. The fish were bulk weighed, counted and their percentage
daily gain and food conversion ratios calculated and compared every
month for three months.
The results indicated that there was no significant difference in fish
growth and performance due to the DNA vaccine. Another study was
also conducted that compared the i.m-delivered IHNV DNA vaccine
when it was administered either jointly or separately with an i.p-
injected polyvalent bacterial oil-adjuvanted vaccine using a similar
experimental design.
This study also examined the concept of antigenic competition by
evaluating the immune response against IHNV when four different
bacterial antigens in an adjuvant were simultaneously delivered. The
results again indicated that there was no significant difference in fish
growth and performance due to the DNA vaccine, polyvalent bacterial
oil-adjuvanted vaccine, or when the two vaccines were used together.
BACTERIAL PATHOGENS
Flavobacterium psychrophilum, the aetiological agent of coldwater
disease and rainbow trout fry syndrome, has emerged as one of
the most significant bacterial pathogens in salmonid aquaculture
6 AQUACULTURE HEALTH INTERNATIONAL NOVEMBER 2006
THE CLEAR SPRINGS FOODS RESEARCH
AND DEVELOPMENT TEAM
worldwide. Previous studies with Dr Ken Cain and his graduate
student, Ben LaFrentz (University of Idaho, Moscow, Idaho), have
suggested that the O-polysaccharide (O-PS) component of the
lipopolysaccharide (LPS) of F psychrophilum is highly immunogenic,
and may be involved in eliciting a protective immune response in
rainbow trout.
Further characterisation of the carbohydrate antigens of
F psychrophilum by SDS-PAGE and western blotting revealed the
presence of two distinct carbohydrate banding patterns. One banding
pattern corresponds with the LPS, and we hypothesise that the other
carbohydrate banding pattern is the loosely associated slime layer, or
glycocalyx, of F psychrophilum.
Electron microscopy of F psychrophilum cells immuno-gold labeled
with a monoclonal antibody specific for this banding pattern
supports this hypothesis, as the outermost layer of the bacterium was
heavily labeled.
This is a significant finding, because the antigens that have been
referred to as the O-PS of LPS (and implicated as a potential vaccine
candidate antigen) appear to be components of the glycocalyx of
F psychrophilum and not LPS.
Passive immunisation experiments utilising the monoclonal
antibody specific for the glycocalyx are underway to determine if
antibodies to the glycocalyx of F psychrophilum provide protection to
rainbow trout following experimental CWD challenge.
If protection is conferred, future research will focus on purification
and further characterisation of the glycocalyx as a potential vaccine
candidate antigen for CWD and RTFS.
CONCLUSION
In summary, Clear Springs Foods maintains an active in-house
service and research programme, along with collaborating with a
number of different research groups on a wide variety of projects.
Increased market competitiveness will only occur through reducing
production costs. Research in the areas of feed and ingredients is
required to enhance fish performance and identify acceptable and
least-cost fish meal and fish oil replacements.
Tremendous gains in production efficiencies could be realised
through rainbow trout strain improvement for growth and disease
resistance. Biosecurity will continue to be a critical feature in any
fish health management strategy. Because antibiotics will be relied
upon less and less, there is a huge need for vaccines and vaccine
delivery strategies, along with identifying potential feed additives that
could enhance innate immune defense mechanisms. These are our
challenges for the future. ■

BACTERIAL HAEMORRHAGIC
SEPTICAEMIA IN TILAPIAS
BY GINA CONROY AND PROFESSOR DAVID A CONROY (PHARMA-FISH SRL, VENEZUELA)
B
acterial haemorrhagic septicaemia, or BHS, has become
recognised as being one of the most common disease
problems that occur in tilapia farming operations on a
worldwide basis.
This condition can be associated with the presence of several
different types of bacteria that produce similar clinical signs in
fish, and for this reason it is preferable to speak of the problem as
the bacterial haemorrhagic septicaemia syndrome.
Tilapias affected by BHS generally show quite characteristic
external and internal clinical signs. The fish tend to be darker in
colour, lethargic and anorexic. One or both eyes may be swollen, a
condition known as exophthalmos.
The most common external signs include areas of haemorrhage,
frequently accompanied by varying degrees of ulceration and
necrosis on the body surface, at the bases of the pectoral and
ventral fins, and in the eye region. (See Figure 1)
Internally, haemorrhagic foci are commonly observed in the
heart, liver, kidney and spleen, and also on the surface of the
viscera and the abdominal cavity. Necrosis is usually present in the
heart, liver, spleen and skeletal muscle tissue, and particularly in
the haematopoietic tissue of the head kidney (see Figure 1). As the
name of the syndrome suggests, outbreaks are always characterised
by the presence of septicaemia, and both free and macrophage-
ingested bacteria can be detected in stained blood films obtained
from diseased tilapias.
BHS is capable of producing mortalities ranging from five to 100
percent in tilapias farmed in fresh and salt water environments.
Bacterial types isolated from outbreaks of BHS in tilapias include
the following Gram-negative rods: Aeromonas hydrophila and
other motile aeromonads (= the “Aeromonas hydrophila complex”,
FOCUS ON FINFISH
FIGURE 1: EXTERNAL
ULCERATION AND
NECROSIS IS EVIDENT
IN THESE EXAMPLES OF
BHS-AFFECTED FISH. NOTE
ALSO THE NECROSIS AND
HAEMORRHAGIC FOCI
PRESENT IN THE INTERNAL
ORGANS OF A TILAPIA
causing motile Aeromonas septicaemia, MAS), Edwardsiella tarda,
Pasteurella multocida, Pseudomonas fluorescens and Vibrio spp.
The example of BHS associated with Vibrio spp is of particular
interest, as cases of vibriosis can occur in both fresh and salt water.
Plumb (1997) reported that Vibrio anguillarum, V parahaemolyticus
and V vulnificus are more frequently isolated from outbreaks
occurring in salt water, whereas V cholerae, V mimicus and
V parahaemolyticus are more common in outbreaks in fresh water.
Edwardsiella tarda is of considerable practical importance, as
this organism is capable of producing cavities in the muscle tissue
which become filled with hydrogen sulphide gas, thereby seriously
affecting the appearance of the fillets, and constituting a problem
which may require the processing plant to be temporarily shut
down for thorough cleansing and disinfection.
Certain of the Gram-positive streptococci (eg Streptococcus
agalactiae, S iniae) are also capable of producing clinical signs of
BHS in cases of acute infections.
It is of the greatest importance that outbreaks of BHS should
be correctly diagnosed as soon as the first cases are detected
on site. The diagnostic procedures must include isolation and
identification of the possible aetiological agent in a bacteriology
laboratory. For this, representative samples must be taken from the
affected organs - using suitable bacteriological techniques - and
streaked onto plates of trypticase soy agar or blood agar containing
five percent sheep blood cells.
Once pure cultures have been obtained, these must be identified
to generic and specific levels on a basis of suitable phenotypical
typing methods in the laboratory. Table 1 outlines certain basic
characteristics that differentiate the various types of Gram-
negative organisms most frequently associated with outbreaks
NOVEMBER 2006 AQUACULTURE HEALTH INTERNATIONAL 7
FOCUS ON FINFISH
BACTERIAL HAEMORRHAGIC SEPTICAEMIA IN TILAPIAS
TABLE 1. BASIC DIFFERENTIAL CHARACTERISTICS OF IMPORTANT GRAM NEGATIVE BACTERIA ASSOCIATED WITH BHS IN TILAPIAS.

TEST REACTION
AEROMONAS HYDROPHILA EDWARDSIELLA TARDA PSEUDOMONAS FLUORESCENS VIBRIO SPP.
GRAM STAIN — — — —
MORPHOLOGY ROD ROD ROD ROD
MOTILITY + + + +
CYTOCHROME OXIDASE + — + +
O/F OF GLUCOSE IN HUGH & LEIFSON MEDIUM F(+ GAS) F O F(NO GAS)
H2S PRODUCTION . + — —
SENSITIVITY TO O/129 VIBRIOSTAT (150 U/DISC) — — — +
PRODUCTION OF FLUORESCENT PIGMENT — — + —

of BHS in tilapias.
Most of the bacteria involved in cases of BHS are normal
components of the bacterial flora of the fish themselves, and of
the aquatic environment in which they are being farmed. It follows
from this that prevention must be based on good management
practices throughout the entire production process.
The water and soil quality must be constantly monitored and
maintained at optimum levels (eg with particular reference to
dissolved oxygen, temperature, pH, hardness, ammonia, nitrite and
other physico-chemical parameters in the case of the water).
Care must be exercised to ensure that stocking densities are kept
well within the limits established for the corresponding stages of
growth of the fish, and great attention must be paid to the use of
suitable feeds and correct feeding practices.
Information has become available on the successful use of
suitable vaccination techniques as a means of preventing massive
outbreaks of edwardsiellosis and Aeromonas hydrophila-associated
BHS in tilapia farming activities.
The application of antimicrobial compounds in the treatment of
clinical cases of BHS must only be contemplated in cases where the
aetiological agent has been duly isolated and correctly identified in
the laboratory, and the minimum inhibitory concentration of the
antimicrobial compound whose use is contemplated has been duly
established and pre-determined on an individual basis.
The administration of any type of antimicrobial compound
should only be undertaken on the basis of the considered
professional advice of the fish health specialist responsible for the
disease status of the tilapia farm concerned, and the treatment
schedule must be carried out under the direct supervision of that
specialist.
The ad libitum use of antimicrobials without such professional
advice is not recommended, as this practice could lead to the
development of antimicrobial-resistant strains of the bacteria,
and would complicate matters further in the event that future
outbreaks of BHS involving antimicrobial-resistant strains of the
same types of bacteria should occur.
REFERENCE
Plumb JA 1997. Infectious Diseases of Tilapia. In: Tilapia
Aquaculture in the Americas (Editors: BA Costa-Pierce and JE
Rakocy), Vol. 1, World Aquaculture Society, Baton Rouge, USA.
pp212-228 ■

ATLAS OF TILAPIA HISTOLOGY 2006

BOOK REVIEW

By CM Morrison, K Fitzsimmons and JR Wright. The World Aquaculture

Society, Louisiana, USA. 1st edition.
BY PROFESSOR DAVID A CONROY, PHARMA-FISH SRL, VENEZUELA

T
ilapia farming is currently one of the most widespread and addition to other
successful types of warm-water aquaculture in the world. In workers in the field
spite of the significant advances made with respect to several of tilapia farming.
aspects of the biology and culture of tilapias, a few “gaps” still remain The individual
in relation to our knowledge of these fish. organs and systems
The recent appearance of the Atlas of Tilapia Histology, authored are dealt with in a
by Drs Carol Morrison, Kevin Fitzsimmons and James Wright Jr, and methodical manner,
published by the World Aquaculture Society, has very successfully and will enable
filled one such “gap”, the normal histology of the tilapias. possible “pathological or other such
In the 96-page document, with abundant full-colour illustrations alterations” to be interpreted in the light of what is “normal”. This
and detailed explanatory notes, the authors describe the anatomy publication should be at the fingertips of all those who have any
and histology of the tilapias, making special reference to the Nile need to examine and interpret histological sections obtained from
tilapia (Oreochromis niloticus). tilapia material.
The histological descriptions are extremely detailed and clearly The authors, as the publishers, are to be congratulated on making
illustrated, making this document an important, original and this important document available. It addresses a definite need in
authoritative source of reference on the normal histology of tilapias respect to the biology and culture of tilapias in general, and can be
for biologists, veterinarians and other fish health specialists, in fully recommended. ■

8 AQUACULTURE HEALTH INTERNATIONAL NOVEMBER 2006

A N OV E R V I E W

STREPTOCOCCUS
IN WARM-WATER FISH
DR JOYCE J EVANS, DR PHILLIP H KLESIUS, DR CRAIG A SHOEMAKER
USDA, ARS, AQUATIC ANIMAL HEALTH RESEARCH LABORATORY, USA

This overview will focus on the epidemiological aspects of Streptococcus iniae and
S agalactiae in warm-water fish, including global distribution, fish host susceptibility, disease
signs, sample collection, transport and storage for successful survival and isolation,
identification by conventional, automated and molecular diagnostic techniques, antibiotic
sensitivities, and vaccination and environmental influences on disease susceptibility.
Overall, this article will chronicle the road from the emergence of these piscine pathogens
to detection, understanding, treatment and prevention.

D
espite being known mainly as mammalian disease agents, Streptococcus agalactiae is the only streptococcal species that has a
Streptococcus agalactiae and S iniae have become recognised group B antigen.
as important pathogens of wild and cultured fish, causing Despite this, most of the non-haemolytic GBS fish isolates were
severe economic losses to the aquaculture and fisheries industries. originally unspeciated, Robinson and Meyer 1966, Plumb et al 1974,
Although S agalactiae has its origins in terrestrial mammals, whereas Baya et al 1990, or misidentified as Streptococcus difficile, Eldar et al
S iniae originated from aquatic mammals, their zoonotic potential 1994.
has not been adequately explored. Streptococcus agalactiae, Lehmann Many of these isolates have now been characterised as Group
and Neumann 1896, or Lancefield’s group B streptococcus GBS B S agalactiae, Wilkinson et al 1973, Elliott et al 1990, Vandamme
has long been recognised as causing bovine mastitis and human et al 1997. Kawamura et al 2005 have proposed a reclassification
neonatal infections, Elliott et al 1990. of S difficilis to unify S difficilis and S agalactiae, with S difficilis
More recently, Evans et al 2006a reported the isolation of regarded as the junior synonym of S agalactiae. Twelve years lapsed
S agalactiae from an aquatic mammal. The first report of GBS before S agalactiae (haemolytic) was again reported in fish kills,
in freshwater fish, golden shiners Notemigonus crysoleucas, Evans et al 2002a.
Robinson and Meyer 1966, preceded that of S iniae by 10 years. Pier and Madin 1976 and Pier et al 1978 first described Streptococcus
iniae from a warm-blooded captive vertebrate, the
Table 1 Dolphin and fish hosts naturally or experimentally infected with Streptococcus iniae, S. agalactiae* freshwater Amazon dolphin, Inia geoffrensis. These S
and both S. iniae and S.agalactiae ‡* from freshwater or estuarine and marine environments.
iniae isolates are the American Type Culture Collection
Freshwater Estuarine/Marine ATTC, Rockville, MD type strains 29178 and 29177,
Amazon dolphin (Inia geoffrensis) Bottlenose dolphin (Tursiops truncatus)*
Cichlidae Ariidae Sciaendiae
and serve as the standard identification of S iniae
Blue tilapia (Oreochromis aureus) Sea catfish (Arius felis)* Red drum (Sciaenops ocellatus) phenotypic characteristics. Eighteen years after its
Mossambique tilapia Carangidae Spot (Leiostomus xanthurus)* initial characterisation, S iniae and the erroneous S
(Oreochromis mossambicus) Yellowtail (Seriola quinqueradita)‡* Atlantic croaker
Tilapia hybrid Clupeidae (Micropogon undulatus)*
shiloi, a junior synonym of S iniae, was reported as the
(Oreochromis niloticus x O. mossambicus)‡* Gulf menhaden (Brevoortia patronus)* Silver trout (Cynoscion nothus)* causative agent responsible for fish kills in cultured
Red tilapia (Oreochromis niloticus x O. Fundulidae Sea trout (Cynoscion regalis)*
aureus)‡* Gulf killifish (Fundulus grandis)* Common mackerel (Scomber japonicus) hybrid tilapia Oreochrimis niloticus x O aureus in the
Nile tilapia (Oreochromis niloticus) ‡* Dasyatidae Jack Mackerel (Trachurus japonicus) United States and Israel, and in cultured trout in Israel,
Tilapia spp. unspecified (Oreochromis spp.)* Stingray (Dasyatis sp.)* Serranidae
Red Tilapia tetrahybrids (Oreochromis Haemulidae Brown spot grouper (Epinephelus coioides) Perera et al 1994, Eldar et al 1994, 1995a.
Grunt (Haemulidae sp.) European sea bass (Dicentrarchus labrax)
mossambicus x O. urolepsis x O. niloticus x O.
aureus )* Black margate (Anisotremus sp.)
Synonyms for S iniae disease derived from the
Centrarchidae Lined piggy (Pomadasys stridens) Siganidae literature include hemolytic streptococcal disease,
Bluegill (Lepomis macrochirus)* Kyphosidae Marbled spinefoot (Siganus rivulatus)
Green sunfish (Lepomis cyanellus)* Bermuda sea chub (Kyphosus sectatrix) Dusky spinefoot (Siganus fuscescens)
bacterial meningoencephalitis, mad fish disease, golf
Cyprinidae Latidae Whitespotted spinefoot (S. canaliculatus) ball disease of freshwater dolphin and S shiloi.
Sheepshead minnow (Cyprinodon variegatus)* Barramundi (Lates calcarifer )
Red-tail Black shark (Epalzeorhynchos bicolor) Lutjanidae Sparidae
Rainbow shark (Epalzeorhynchos erythrurus) Snapper (Ocyurus chrysurus) Gilthead sea bream (Sparus auratus) ‡* GLOBAL DISTRIBUTION
Golden shiners (Notemigonus crysoleucas)* Pinfish (Lagodon rhomboids)* The geographical distribution of Streptococcus iniae
Zebrafish (Danio rerio) Moronidae Stromateidae
Moronidae Striped bass (Morone saxatilis)* Silver pomfret (Pampus argenteus)*
and S agalactiae GBS is primarily confined to temperate
Hybrid striped bass/ Sunshine bass Mugilidae Synodonttidae and tropical regions that culture warm water fish.
(Morone chrysops x M. saxatilis) Borneo grouper (Liza macrolepis) Lizard fish (Synodus variegates)
Mugilidae Klunzingeri mullet (Liza klunzingeri)* These bacterial species have been reported from six
Gray mullet (Mugil cephalus) Striped mullet (Mugil cephalus)* continents and 23 countries. Reports of S iniae have
Plecoglossidae Paralichthyidae been more numerous, occurring in more countries and
Ayu (Plecoglossus altivelis) Japanese/ Olive flounder
Salmonidae (Paralicthys olivaceus)
fish hosts than S agalactiae.
Amago salmon Pomatomidae Group B Streptococcus has been reported from six
(Oncorhynchus rhodurus var. macrostomus) Bluefish (Pomatomus saltatrix)*
Rainbow trout (Oncorhynchus mykiss)‡* Scaridae countries on three continents: United States (North
Coho salmon (Oncorhynchus kisutch) Parrot fish (Sparisoma aurofrenatum)/
Terapontidae
America), Israel, Japan, Kuwait, Thailand (Asia) and
(S. viride)
Jade Perch/Barcoo grunter (Scortum barcoo) Brazil (South America). Countries in which both S
Ictaluridae
Channel catfish (Ictalurus punctatus) ‡*
iniae and S agalactiae have been reported include
the United States (North America), Israel, Japan and

10 AQUACULTURE HEALTH INTERNATIONAL NOVEMBER 2006

Thailand (Asia). (Figure 1) FIGURE 1
Kitao 1993 indicated that 22 species of wild or cultured fish
were susceptible to S iniae. Since that time, the number of species
reported to be susceptible has doubled. More than 50 species in 29
families of freshwater, estuarine and marine fish have been reported
to be susceptible to either S iniae or S agalactiae, indicating a broad
host range and environmental adaptability of these organisms.
(Table 1)
Both S iniae and S agalactiae are more than likely under-
represented geographically and in numbers of fish hosts, due to the
absence of formally published reports or inadequate identification
and speciation.
Streptococcus iniae has been reported in several tilapia species
Oreochromis spp from 14 countries. Three species of tilapia have
been naturally infected with Group B Streptococcus agalactiae in
Israel, Thailand and Brazil. Natural S agalactiae infections in
tilapia cultured in the USA have not been reported. Despite the
initial reports of non-haemolytic GBS causing fish kills in cultured frozen at -20° C for nine months. The use of frozen fish may prove
freshwater golden shiners in the USA and later rainbow trout to be a useful archival or retrospective alternative to fresh fish for
Oncorhychus mykiss in Israel, the majority of natural infections of recovering pathogenic streptococci in instances when diagnostics
S agalactiae in the USA, Japan, Eldar et al 1994, and Kuwait, Evans et are unavailable or impractical.
al 2002a, Duremdez et al 2004 have involved non-tilapine estuarine
and marine fish. The reader is referred to Evans et al 2006b for a IDENTIFICATION
review of S iniae and S agalactiae in tilapia species. There has been significant confusion in the published literature with
respect to which streptococcal species is the agent responsible for
DISEASE SIGNS causing disease signs, mortality and pathology in fish. Identification
Fish infected with either Streptococcus iniae or S agalactiae of S iniae and S agalactiae is hampered by modifications in the
show similar acute neurotropic disease signs. The behavioral taxonomy of the Streptococcus genus, the source of infection and
manifestations of these infections include abnormal, spiral or erratic familiarisation of the diagnostician with other Gram-positive
swimming, swimming and whirling at the surface, and head-up or bacteria that infect fish.
tail-up swimming. The hallmark of streptococcal infection is a C Accurate comparison of an isolates’ phenotypic characteristics
or comma-shaped body curvature. Common ocular abnormalities to Type strains and recognised mammalian bacterial species,
include peri-orbital and intraocular haemorrhage, opacity and
exophthalmia in the chronic stage of infection.
Reddening and haemorrhage are often noted in the integumental
and muscoskeletal system, particularly cranially and around the
mouth, snout, operculum and fins. Darkened coloration is also
commonly noted in infected fish. Anomalies that have not been
reported from S iniae-infected fish but from S agalactiae infections
include an unusual opercular clearing or “window to the gill”, Evans
et al 2002a, and long mucoid faecal cast, Pasnik et al 2005.

SAMPLE COLLECTION
Due to the sudden occurrence of streptococcal disease outbreaks
in remote locations worldwide and the absence of trained fish
diagnosticians or adequate facilities in these areas, fish collection,
storage and transport techniques that can be performed by
aquaculturists are needed.
The determination of proper collection, storage, transport and
isolation techniques for a streptococcal pathogen can enhance the
recovery, characterisation and identification of the pathogen. One
of the first steps in the collection process should be deciding where
the cultures are obtained from the fish.
The brain and anterior kidney are common sites of bacteriologic
culture of Streptococcus iniae, Shoemaker et al 2001. However,
non-lethal bacteriological sampling of the nares may also be useful
during the early stages of S iniae infection, Evans et al 2001.
Evans et al 2002ab demonstrated the survival of S agalactiae
collected from naturally infected fish using dry swabs and in
transport for four days, and survival of S iniae and S agalactiae
for up to 11 days following dry swab sampling and enrichment in
experimentally infected fish.
The length of time tissues can remain frozen and still yield
viable Streptococcus sp has been determined from frozen fish, both
naturally and experimentally infected with S agalactiae, Evans et al
2004a. Streptococcus agalactiae could be recovered from fish tissues

NOVEMBER 2006 AQUACULTURE HEALTH INTERNATIONAL 11

A N OV E R V I E W
STREPTOCOCCUS IN WARM-WATER FISH

Table 2. Conventional tests to confirm Streptococcus genus and discriminate selection of appropriate microbiological media and laboratory tests
between Streptococcus iniae and Streptococcus agalactiae. and correct interpretation of laboratory tests, are important.
Conventional Tests S. iniae S. agalactiae Alterations in the taxonomy and nomenclature of the Streptococcus
Gram stain + + genus have occurred as a result of the advent of molecular
Catalase - -
Hemolysis on 5% sheep blood agar + +/-
techniques employed to help delineate differences in bacterial
Motility - - genera and species. In addition, similarity among the phenotypic
Growth @ 10°C + - characteristics of catalase negative, Gram-positive coccus genera
@ 45°C * - -
in 6.5% NaCl broth - - such as Lactococcus and Enterococcus and haemolytic Streptococcus
Reaction on Bile -Esculin media - - spp can cause confusion and misidentification.
Production of :
Pyrrolidonyl arylamidase (PYR) + - The genus Streptococcus formerly contained both Lactococcus and
Leucine aminopeptidase (LAP) + + Enterococcus. The Streptococcus genus has since been split into three
Gas from glucose in MRS broth - -
Acid in sorbitol - - distinct genera: Streptococcus, Enterococcus and Lactococcus, Facklam
CAMP test + + 2002.
Voges-Proskauer (VP) reaction ‡ - -/+
Susceptibility to Vancomycin (30μg) - - Numerous earlier reports of unspeciated Streptococcus spp are
Hydrolysis of: not Streptococcus species at all, based on the current classification
Hippurate - +
Starch + -
system. For example, in pathogenesis studies conducted in yellowtail
Lancefield group None B with the Japanese YT-3 strain, the isolates were initially referred to as
* Perera et al. (1994) reported atypical S. iniae growth at 45°C; ‡ Commercial multitest
API systems give a positive VP reaction for S. agalactiae.
a Streptococcus sp, Kimura and Kusuda 1982, but are now recognised
as Lactococcus garvieae, formerly Enterococcus seriolicida.
Likewise, the Streptococcus sp in rainbow trout described by
Boomker et al 1979, has been identified as a Lancefield’s group D
Enterococcus. Therefore, researchers should heed the taxonomic
changes in the Streptococcus genus, and be aware that references to
Streptococcus spp in older literature use outdated nomenclature.
The investigator should perform the conventional laboratory
tests in Table 2 to determine the phenotypic characteristics of
the bacteria, confirm the isolate as a member of the Streptococcus
genera, and differentiate between S agalactiae and S iniae.
These include the Gram stain, catalase test, haemolysis on
trypticase soy agar containing five percent sheep blood, motility,
growth at 10° C and 45° C, growth in broth containing 6.5 percent
NaCL, reaction on bile-esculin BE medium, production of leucine
STREPTOCOCCUS INIAE AND S AGALACTIAE DISEASE aminopeptidase LAP, pyrrolydonyl arylamidase PYR, gas from
SIGNS IN NILE TILAPIA: OCULAR DISEASE glucose in Mann, Rogosa, Sharp MRS Lactobacillus broth and
acid in sorbitol broth, CAMP test for enhanced beta haemolysis
production by Streptococcus using Staphylococcus aureus, Voges-
Proskauer reaction VP for production of the enzyme acetymethyl
carbinol, hydrolysis of hippurate and starch, susceptibility to
vancomycin 30µg and Lancefield group.
As part of the Streptococcus genus, S iniae and S agalactiae are
Gram-positive, catalase-negative cocci in pairs or chains. Both are
non-motile, susceptible to vancomycin, negative in the VP reaction,
and do not react on BE media, produce acid in sorbitol, gas in MRS
broth, or grow at 45° C or in broth containing 6.5 percent NaCl,
although some strains may show weak growth.
Both S iniae and S agalactiae are positive for LAP and CAMP.
To confirm the diagnosis of S iniae and differentiate between
S agalactiae, S iniae, unlike S agalactiae, usually grows at 10° C, does
not hydrolyse hippurate but hydrolyses starch, and does not possess
a Lancefield group.
S iniae is always haemolytic on blood agar plates, whereas S agalactiae
can be either haemolytic or nonhaemolytic. Starch hydrolysis testing
is essential for identification of S iniae and differentiation from
other non-starch hydrolysing streptococcal organisms, Evans et al
2004a. Lancefield serological grouping and starch hydrolysis testing,
however, is lacking from many S iniae characterisations.

COMMERCIAL TESTS
Commercial multi-test system kits API 20 Strep, Rapid Strept 32, API
CH 50 and Vitek often used in the rapid presumptive identification
of streptococci should be validated by conventional tests. These
commercial multi-test systems cannot identify S iniae, as this
organism is not in these systems’ databases, Evans et al 2004a.
Furthermore, the biochemical reaction for starch in multi-test
systems API 20 Strep and API CH 50 is acidification, not hydrolysis.
Fish researchers began and continue utilising API multi-test systems

12 AQUACULTURE HEALTH INTERNATIONAL NOVEMBER 2006

to identify S iniae, and many substitute acidification of starch
for starch hydrolysis in publications. Variability in phenotypic
characteristics of Streptococcus species has been reported using these
test systems and may reflect artifacts of the assays. Vandamme et
al 1997 reported differences in hippurate hydrolysis between GBS
using API test systems incubated at different temperatures.
Evans et al 2002a reported positive VP reactions for GBS isolates
in API systems and negative VP reactions by conventional methods.
Excellent results, however, have been obtained using the Biolog
system Hayward CA, USA for characterisation and identification of
S iniae and S agalactiae, although atypical S agalactiae isolates are
not always identified correctly.
MOLECULAR TECHNIQUES
Klesius et al 2006a developed a rapid and sensitive monoclonal-
antibody-based technique for detecting S iniae. Several molecular
techniques have been developed as rapid, sensitive and specific
complements to conventional diagnostic and taxonomic protocols
for S iniae and S agalactiae. The molecular techniques include
randomly amplified polymorphic DNA RAPD, restriction
fragment length polymorphism RFLP, amplified fragment length
polymorphism AFLP and whole genome DNA-DNA hybridisation,
though polymerase chain reaction PCR has been the most widely
studied.
Specific PCR primer sequences obtained from genes or DNA
fragments have been successfully used to identify S iniae and
S agalactiae infections in fish, Berridge et al 2001, Mata et al 2004,
Kawamura et al 2005.
PCR targets in these studies have included the 16S rRNA, 23S
rRNA, 16S-23S RNA intergenic spacer, DNA gyrase, superoxide
dismutase, and lactate oxidase genes. PCR and whole-genome
DNA-DNA hybridisation were used to identify S difficile as a group
B Streptococcus sp, S agalactiae, based on genetic homogeneity,
Vandamme et al 1997, Berridge et al 2001, Kawamura et al
2005. However, molecular techniques have not always successfully
distinguished between strains that exhibited biochemical differences,
Dodson et al 1999.
ANTIBIOTIC SENSITIVITIES
Streptococcus agalactiae and S iniae are sensitive to numerous
antibiotics including oxytetracycline, ciprofloxacin, amoxicillin-
clavulanic acid, penicillin, chloramphenicol, tetracycline, rifampin,
sulfamethoxazole trimethoprim, erythromycin and vancomycin.
Streptococcus iniae isolates are resistant to streptomycin, amikacin,
nalidixic acid and colistin, and sensitivities vary to gentamicin and
ampicillin, Pier and Madin 1976, Eldar et al 1994, Perera et al 1994.
The sensitivities of GBS fish isolates to bacitracin, gentamicin,
streptomycin and novobiocin have varied, Baya et al 1990, Wilkinson
et al 1973, Evans et al 2002a. Israeli GBS isolates were resistant
to gentamicin, amikacin, nalidixic acid, colistin, novobiocin and
optochin, Eldar et al 1994, Vandamme et al 1997.
DISEASE SUSCEPTIBILITY
Vaccination is an alternative to chemotherapeutics. The reader
is referred to Klesius et al 2006b for a review of streptococcal
vaccinology in warm-water fish. Currently, efficacious S agalactiae
and S iniae extracellular product ECP vaccines have been developed
and patented. (US patent # 0208077 A1 and # 6,379,677 B1.)
These vaccines produce antibody-mediated acquired immunity
and therapeutic effects, Shelby et al 2002, Pasnik et al 2005, Evans
et al 2006c. The potential of stress and vaccination, Evans et al
2004c, and poor water quality, such as sub-lethal dissolved oxygen,
Evans et al 2003, for causing increased susceptibility to S agalactiae
disease has been experimentally evaluated in Nile tilapia using a
simple blood glucose monitor. This research has demonstrated
the link between disease and low dissolved oxygen stress, and the
protective effect of the vaccination process on disease resistance.
In addition, this technology may be possibly amenable to field
assessment of fish stress at aquaculture facilities and in the wild, and
is of practical application to fish farmers to mitigate streptococcal
disease outbreaks.
Conversely, research has demonstrated that exposure to
toxic unionised ammonia may not increase S agalactiae disease
susceptibility as previously believed, Evans et al 2006d. Chang and
Plumb 1996 observed increased susceptibility of injured Nile tilapia
to S agalactiae at elevated salinities at 25˚ C and 30˚ C. Shoemaker et
al 2000 evaluated the practical considerations of cultured Nile tilapia
density and mortality rates following immersion or cohabitation
exposure to S iniae. Tilapia densities of 11.2 g/l and above had a
significant affect on S iniae mortality.
SUMMARY
Streptococcus agalactiae and S iniae are clearly important fish
pathogens. It is unclear if their emergence as fish pathogens is
associated with heightened recognition of their occurrence in
recent fish kills. It is likely that their existence has been previously
under-reported by the fish health community. The advent of
increased awareness and visibility of these pathogens and adequate
confirmatory diagnostic techniques may enhance their frequency
of detection worldwide. Epidemiological considerations as to the
source, origin, potential for zoonotic infection by streptococcal
organisms, and effective treatment and prevention against these
organisms emphasise the importance of accurate identification.
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A N OV E R V I E W
STREPTOCOCCUS IN WARM-WATER FISH
BODY AND FIN HAEMMORHAGIC LESIONS IN
FISH INFECTED WITH STREPTOCOCCUS
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new NODAVIRUS FOUND IN
NORWEGIAN FARMED COD
Jan Fredrik Frantzen, Fiskeriforskning, Norway
Earlier this year, the National Veterinary Institute and the
Norwegian Food Safety Authority reported that nodavirus,
which has previously affected both halibut and turbot fry in
Norway, has been detected for the first time in Norwegian
farmed cod. The virus can cause the disease viral nervous
necrosis (VNN) in fish that become infected. The result can
be reduced appetite and high mortality.
Senior scientist Ann-Inger Sommer, engineer Saskia Mennen
and others have spent several years studying infection by
nodavirus in halibut, spotted wolf fish and cod. Sommer says
that in cod, as in the other fish species, fry are most vulnerable
to VNN after infection through the water.
Tests have shown that the smallest cod fry become sick
after infection with a nodavirus from Norwegian farmed
cod. During the two-month test period, 56 percent of the
cod fry died that weighed 0.5g when infected via the water.
Conversely, larger fry that weighed 5g did not die of infection
via the water. In this group, 35 percent died of VNN when they
were infected through injection of the virus into the fish.
The last tests also show that when the temperature in the
water rises, there is a pronounced increase in mortality in fish
infected with the virus. Fry weighing 5g infected by injection of
the virus suffered 60 percent mortality in water kept at around
15˚C. The mortality rate was much lower in cooler water.
“The tests show that infected cod are far more likely to die of
nodavirus if the water temperature rises”, says Sommer.
The research results at Fiskeriforskning are the beginning of
a challenge model for the disease VNN in cod, on a level with
the model scientists at the institute have created for the viral
disease IPN in salmon. For example, such a challenge model
can help scientists create a vaccine against VNN in cod, or
used in the cod-breeding programme to select cod families
that are most resistant to the disease. “When complete, the
model will also help us learn how cod thrives best in farming
conditions,” says Sommer. “A fish that that thrives also has a
better immune defence.”
The Research Council of Norway, Innovation Norway and
Fiskeriforskning financed the research.
Contact the following Fiskeriforskning staff:
Senior scientist Ann-Inger Sommer (+47) 77 62 92 01
Director of Research, Erling Sandsdalen (+47) 77 62 92 17
Photo caption: Atlantic cod (Courtesy of Frank Gregersen,
Fiskeriforskning)
AN ATLANTIC COD
(PICTURE COURTESY OF
FISKERIFORSKNING)
16 AQUACULTURE HEALTH INTERNATIONAL NOVEMBER 2006
FISH AND DNA CHIPS
(Source: Tracey Duncombe, BBSRC and Professor Alan Teale,
University of Stirling)
Salmon farmers and wildlife biologists share a common
dilemma: How do you assess well-being in an animal that
often gives few indications that things are badly wrong before
being found dead?
Research by the Exploiting Genomics Initiative at the
Biotechnology and Biological Sciences Research Council is
being used to develop a genetic tool that will make it easy
to monitor the health and performance of Atlantic salmon.
The work is set to benefit the salmon farming industry,
and could also help conserve and restock wild salmon
populations.
Collaborative research between scientists at the
Universities of Stirling, Aberdeen and Cardiff, together with
ARK Genomics at the Roslin Institute and researchers at the
Norwegian School of Veterinary Science is developing a DNA
chip to monitor indicators of health and performance.
The Atlantic salmon Salmon salar is the most important
farmed fish species in the United Kingdom, and a
disappearing species in the wild. The idea behind the
DNA chip is to take a “snapshot” of indicator biological
processes within the fish, from small tissue samples. It is the
culmination of a four-year, multi-centre study to identify
the genetic basis of commercially important traits. The
project, known as Salmon TRAITS (TRanscriptome Analysis
of Important Traits in Salmon), involves a number of
industrial partners: Operon Biotechnologies Gmbh, Marine
Harvest and Scottish Quality Salmon, an organisation that
represents the producers of 65 percent of Scottish farmed
salmon.
“The four most important constraints on commercial
production are the supply of contaminant-free highly
unsaturated oils, including omega-3s, for the salmon diet;
the long and complex lifecycle; infectious disease; and
protein growth efficiency,” says Professor Alan Teale, lead
researcher at the Institute of Aquaculture at the University
of Stirling.
“We have identified genes involved in polyunsaturated
fatty acid metabolism, protein metabolism, bacterial and
viral infection, and freshwater-to-seawater adaptation.
Greater knowledge of these processes will help us monitor
the health, development and productivity of farmed and
wild Atlantic salmon populations with a depth and precision
that has not been possible previously,” Prof Teale said.
The life cycle of the Atlantic salmon includes a number of
dramatic physical and body chemistry changes that prepare the
fish for the shift from a freshwater to a saltwater environment.
In nature, this metamorphosis, known as smoltification,
occurs during spring, and marks the start of a period of
intense growth.
However, as little is known about the genetic control of
smoltification, salmon farmers have had to rely on their own
experience to predict the best time to move fish into saltwater
cages. Get it wrong and the consequences can be serious.
It is difficult to tell when fish are ready to be moved.
Farmers have to make a judgement based mainly on the
appearance of the fish. “Move them too early and the fish
may not thrive, becoming stunted and eventually dying, while
stress associated with premature transfer may leave them
more vulnerable to infection,” says Dr Glen Sweeney of the
University of Cardiff.
The Cardiff team has identified a number of genes that
appear to be up-regulated during smoltification in tissues
such as the gill, brain, pituitary and kidney. “The identification
of these markers could be useful in determining the readiness
of fish for saltwater transfer, and would also provide a
means of studying the effects of environmental pollutants on
smoltification,” says Dr Sweeney.
Highly unsaturated fatty acids (HUFA), including omega-3s,
are essential nutrients, important for growth and development
in all animals, particularly the normal functioning of cell
membranes of the central nervous system. However, many
species cannot produce active forms of these oils from plant
oil sources effectively. They need ready-made supplies.
For man, this is why oily marine fish are an important
part of the diet, as wild salmon get most of their omega-
3s through the marine food chain. “The problem is that
feeding fish to salmon in order to supply them with omega-
3s is no longer sustainable, and farmed salmon must
become vegetarian for at least part of the production cycle,”
says Prof Teale.
Research in Stirling is focussed on the genes controlling
synthesis of HUFA in salmon. “Being able to monitor the
function of these genes in fish stocks will enable precision in
dietary formulation and optimisation of feeding levels,” says
Prof Teale. “It will also enable breeders to select for fish in
which these genes are more active than in the majority of the
population.”
Studies of wild salmon populations may also prove
revealing, as HUFAs help nerve cells function across a wide
temperature range. “With climate change, freshwater streams
where salmon parr live are getting warmer,” he says. “But after
smolting, when much of the UK’s wild salmon population
moves into the North Atlantic and out to the Labrador Sea,
they are faced with increasingly lower temperatures. We will
not know until we do the research, but it is possible that these
opposing natural selecting forces partly explain why wild
stocks are in serious decline.”
Given the environment in which farmed fish are maintained,
together with high stocking densities and handling issues,
bacterial and viral pathogens, including a number of emerging
diseases, pose a difficult challenge for aquaculture. Poor health
can often be hard to see. The first indication of a problem may
be large numbers of dead fish.
Professor Secombes’ group at the University of Aberdeen
has analysed gene expression in fish tissue samples, following
infection with different pathogens. Several hundred genes
were found to be increased or decreased following a bacterial
infection in salmon.
“These genes have different roles in the immune response,
including bactericidal activity, acute phase response proteins
and cytokines,” says Prof Secombes. “We have also studied the
relationship between nutritional status and immune response,
and there are indications that the progression of an immune
response may be affected by nutritional status.”
With the large-scale gene expression analysis completed and
performance indicators selected, the final stage of the project
is to construct the DNA chip and confirm its suitability for
use in the aquaculture industry and for wild salmon stocks.
Contact:
Prof Alan Teale, email a.j.teale@stir.ac.uk
Prof Chris Secombes, c.secombes@abdn.ac.uk
Dr Glen Sweeney, SweeneyGE@Cardiff.ac.uk
WINTER ULCER DISEASE
APPEARING IN FRY IN NORWAY
Irene Andreassen (Fiskeriforskning, Norway)
Adding seawater to salmon hatcheries before the fish are
physiologically ready may be contributing to an increase in
the bacterial disease winter ulcer in Norwegian salmon, where
WINTER ULCER DISEASE IN
SALMON (COURTESY OF
FISKERIFORSKNING)
the disease has caused substantial economic losses.
The winter ulcer bacteria, Moritella viscose, is well adapted
to low temperatures both in seawater and brackish water.
If the water is cold, ie below 10˚C, there is an increased
probability of a winter ulcer outbreak.
Tests show that salmon smolts introduced to brackish water
before they are ready will become weakened and their health
compromised. They also exhibit impaired appetite and show
minimal weight gain.
Salmon hatcheries with intensive production which
have low water exchange and add too much seawater will
have favourable environmental conditions for the bacteria.
However, these conditions are unfavourable for the smolts,
and can reduce their resistance to diseases.
“We examined what effect the water quality in the tanks
has for the susceptibility of the smolt to infection with the
winter ulcers bacteria after it is introduced into the sea,” says
a scientist with Fiskeriforskning, Dr Helene Mikkelsen. “We
tested different water qualities such as high and low water
exchange and fresh versus brackish water with two percent
salinity.
“Our results show that mortality after infection was highest
in smolts that had been exposed to brackish water with two news
percent salinity. In contrast, fish that had received optimal
water quality had the lowest mortality,” Dr Mikkelsen
said. “Unfortunately, there is no obligation to report this
disease, and thus it isn’t possible to know the extent of the
problem.”
In Norway, all seawater brought into salmon hatcheries
must be UV-treated to reduce the level of bacteria in the
water, but the treatment is probably not effective enough.
The bacteria that cause winter ulcers most likely has a greater
ability to survive UV radiation than other bacteria naturally
found in seawater.
Fish with winter ulcers are treated by administering in-feed
antibiotics. Indeed, a large part of today’s use of antibiotics in
Norway is due to winter ulcers. “The efficacy of the antibiotic
treatment varies because sick fish have lower appetites and
don’t eat enough of the antibiotic feed. Better control of the
incidence of the disease will thereby also result in less use of
antibiotics,” says Mikkelsen.
The experimental work is part of a larger joint project
between Fiskeriforskning, NIVA, the Norwegian University of
Life Sciences at Ås, the University of Bergen, Marine Harvest
and AquaNet. The project is financed by the Research Council
of Norway, Marine Harvest, AquaNet and Fiskeriforskning.
Contact Dr Helene Mikkelsen (+47) 77 62 90 55
NOVEMBER 2006 AQUACULTURE HEALTH INTERNATIONAL 17
new INTERNATIONAL:
CONFERENCE RAISES AWARENESS
The first Global Conference on Aquatic Animal Health was
dedicated to reinforcing the commitment of OIE-member
countries to their rights and obligations regarding disease
notification and implementation of OIE standards. The
conference was organised by the World Organisation for
Animal Health (OIE), in collaboration with the Norwegian
government,
The OIE director-general, Dr Bernard Vallat, said about
half global fish production came from farming, and that bad
animal health conditions were one of the major constraints to
its development.
“The conference applied itself to raising awareness and
defining roles and responsibilities of national veterinary
services, other competent authorities possibly involved and
the private sector in the prevention and control of aquatic
animal diseases,” Dr Vallat said.
The conference, held in Bergen, Norway from October 9 to
12, established a set of recommendations to create a framework
for improved cooperation among stakeholders to improve
global aquatic animal health monitoring, information sharing
and control of aquatic animal health and welfare issues.
Delegates from the OIE member countries, representatives of
national veterinary services, other competent authorities and
aquaculture producers made up the 150 participants.
The market for ornamental fish is
significant, with an estimated 3.5
million homes in the United Kingdom
The conference served to underpin the efforts of the OIE
and those of the OIE Aquatic Animals Commission chaired
by Dr Eva-Maria Bernoth in promoting training at farm level
up to the veterinary network and authorities level in early
detection and appropriate rapid response to aquatic animal
outbreaks.
It emphasised the need for enhanced information sharing
and official notification in the field of aquatic animal health,
based on the success of the OIE World Animal Health
Information System, or WAHIS, in monitoring and controlling
the occurrence and spread of terrestrial animal diseases.
The OIE also updated the advancement of the elaboration
of scientifically based guidelines on aquatic animal welfare to
be prepared for discussion with all OIE member countries.
These include chapters on live transport of fish, killing for
human consumption and sanitary killing of fish. Similar
chapters concerning terrestrial animals already exist in the
OIE Terrestrial Animal Health Code for Terrestrial Animals.
The OIE, with the support of the Aquatic Animals
Commission, will respond to the outcomes of this first
global conference, and implement these newly outlined
recommendations using its network, including its regional
and sub-regional offices.
USA/CANADA: DISTRIBUTION
AGREEMENT FOR NORTH AMERICA
Eka Chemicals Inc. and Aquatic Life Sciences Inc have
appointed the Aquatic Life Sciences companies Western
Chemical Inc and Syndel Laboratories Ltd as exclusive
distributors for 35 percent PEROX-AID throughout the
United States and Canada.
Eka Chemicals Inc, a leading hydrogen peroxide
18 AQUACULTURE HEALTH INTERNATIONAL NOVEMBER 2006
manufacturer, has been pursuing FDA approval for 35
percent PEROX-AID (35 percent aquatic-grade hydrogen
peroxide) for use on freshwater-reared finfish and their
eggs, in collaboration with scientists at the Upper Midwest
Environmental Sciences Centre and the National Aquaculture
NADA Coordinator. FDA approval is expected in the coming
months for three new label claims. Once approved, the
current low regulatory priority status of hydrogen peroxide
will be rescinded, and 35 percent PEROX-AID will be the
only FDA-approved hydrogen peroxide product for use on
freshwater-reared finfish and their eggs.
“With the pending approval of 35 percent PEROX-AID(r)
we needed a partner with the expertise and distribution
channels to rapidly and effectively distribute our product,”
said the director of Business Development at Eka Chemicals,
Anthony R Colasurdo. “The Aquatic Life Sciences companies
of Western Chemical and Syndel Laboratories are the logical
choice, as they are the leaders in providing fish health
products in the United States, Canada and the rest of the
world.”
Aquatic Life Sciences said it welcomed the opportunity to
incorporate 35 percent PEROX-AID into its line of approved
fish health products, said the chief executive officer, Steven J
Becker. The product would be a valuable and much-needed
tool for its aquaculture customers. “We applaud the work
that Eka Chemicals has put into this approval, and we look
forward to working with them.”
Contact
In the USA: Western Chemical,
Ferndale WA 98248 USA, (+1) 800-283-5292
In Canada: Syndel Laboratories Ltd,
Vancouver BC (+1) 800-663-2282
UK/USA:
KHV VACCINE WORK PROGRESSES
Henderson Morley says it is ready to start field trials of a
candidate vaccine against the koi carp herpes virus. The trials
follow 10 months of work in collaboration with the Hagerman
Aquaculture Research Institute, based in Idaho, USA, under
the supervision of the institute’s director, Professor Ron
Hardy.
The directors of Henderson Morley believe that these
initial immunogenicity studies will produce results within
six months. That should enable Henderson Morley scientists
to optimise the vaccine formulation with a view to securing
a product licence from the veterinary authorities as soon as
practicable.
The board says it is aware that the time taken to commercialise
human products may be significant, so products in the
Henderson Morley portfolio targeted at animal health may
offer the prospect of nearer-term revenue opportunities.
The market for ornamental fish is significant, with an
estimated 3.5 million homes in the United Kingdom and 139
million fish in 13 million homes in the United States, with
koi carp species being among the most popular and most
valuable.
In 1998, widespread outbreaks of mortality occurred in
both fish farms and ornamental ponds, when 85 percent to
100 percent of infected fish died within a few days. It was
subsequently discovered that this disease was caused by a
newly isolated virus - koi herpes virus, or KHV. This virus has
now been isolated in at least 28 countries, including the USA,
Japan and the UK, and has become a significant problem
for koi breeders and enthusiasts. Once infected, a pond will
lose the majority of its fish within days, and no treatment or
vaccine is currently licensed in the UK.
 The impact of KHV is very significant in the koi keeping and CHARITY: AQUACULTURE
breeding community. The National Fisheries Laboratory, in WITHOUT FRONTIERS SEEKS HELP
its notes on koi herpes virus provided by the UK Environment Dr Scott Peddie (Patterson Peddie Consulting Ltd, UK)
Agency, says that, “Due to the pathogenicity of the virus
and difficulties with detection, the Environment Agency is
very concerned about the potential impact of KHV to carp
fisheries within England and Wales.”
The company has appointed Professor Ronald John Roberts,
Professor Emeritus at the University of Sterling, to its Scientific
Advisory Board to help develop the KHV vaccine.
“We are very pleased to welcome Professor Roberts to the
board,” said the chairman of Henderson Morley, Andrew
Knight. “As we concentrate on our animal health research
programme, his vast experience will be of enormous benefit
to the company and its scientific advisory board.”

US LIMITS FISH MOVEMENTS

The United States government issued a federal order on
October 24 immediately prohibiting the interstate transport
AQUACULTURE IS A SMALL-SCALE FAMILY AFFAIR IN THE
of about 37 live fish species from eight states surrounding the AREAS IN WHICH AWF OPERATES
Great Lakes region and two Canadian provinces.
Its purpose was to prevent an outbreak of viral haemorrhagic As many readers of Aquaculture
septicemia, or VHS, from spreading into aquaculture Health International may be
facilities. aware, the independent non-
The order applied to Illinois, Indiana, Michigan, Minnesota, profit organisation Aquaculture
New York, Ohio, Pennsylvania and Wisconsin, and the without Frontiers promotes and
Canadian provinces of Ontario and Quebec. It was distributed supports responsible and sustain-
by the deputy administrator of the United States’ Department able aquaculture initiatives.
of Agriculture’s Animal Plant Health Inspection Service, This in turn helps to alleviate
Veterinary Services. The order was issued pursuant to the poverty by improving rural liveli-
Animal Health Protection Act. hoods in developing and transi-
The list may be amended if other species are identified as tion countries. Formed in 2004,
being susceptible to VHS. AwF is registered as a charity
See www.aphis.usda.gov/vs/aqua in the United Kingdom and as
a non-profit organisation in the
VACINE GAINS United States of America. The
PROVISIONAL AUTHORISATION organisation draws on the expe-
Schering-Plough Animal Health has gained a provisional rience of respected professionals
marketing authorisation for the introduction of AquaVac™ from every relevant discipline,
Relera™, a new bivalent vaccine providing further protection and already has a database of TILAPIA GROW WELL
IN AN AWF-VOSD PROJECT
against enteric redmouth disease (ERM). The immersion more than 100 volunteers located IN BANGLADESH
vaccination allows farmers to enhance existing vaccination across the globe. news
programmes by combining the typical endemic Hagerman AwF is said to be unique in devoting all of its resources
Type 1 ERM-causing strain of Yersinia ruckeri with a new and attention to aquaculture. It does not seek to promote
biogroup called EX5. aquaculture in isolation, but as a component of integrated
EX5 has been responsible for widespread losses in the rural and coastal development plans, and of strategies to
UK rainbow trout industry and appears to be becoming alleviate poverty.
increasingly prevalent. During the past five years the industry For example, AwF is currently teaching very poor families in
has become aware of this new biogroup and the potential for India and Bangladesh how to raise fish for food and income,
it to be as equally destructive as the widespread Hagerman while new projects have been commissioned in Thailand
Type 1 strain of ERM. and Kenya. Its volunteers have also been assisting tsunami
Highlighting the potential for trout farmers, Dr Chris victims in Aceh, Indonesia, to re-start growing fish, shrimp
Gould, global technical services manager at Schering-Plough and seaweed after the loss of their facilities and, in many cases,
Animal Health says, “Existing licensed vaccines do not members of their families.
confer sufficient protection against this new biogroup, and See www.aquaculturewithoutfrontiers.org. Please consider
it has been necessary to develop a new vaccine which offers supporting AwF in any way you can. Instructions are on the
full protection. Through collaboration with the UK trout website.
industry, Schering-Plough Animal Health has been able to
quickly respond to industry needs, isolate the new biogroup INTERNATIONAL:
and develop this new vaccine.” INTERVET SUPPORTS AQUACULTURE
AquaVac™ Relera™ will be available as a POM-V WITHOUT FRONTIERS AND OTHER NGO’S
veterinary product in the UK. The company is supporting the Intervet International B.V. (www.intervet.com) is dedicating
announcement with a series of communications designed to its 2006 Christmas donation (in lieu of sending out Christmas
inform fish vets on the development of AquaVac™ Relera™ and New Year cards) to four non-governmental organisations
and the most appropriate vaccine programmes for clients (NGOs). Four initiatives from different parts of the world
with ERM. have been selected.

NOVEMBER 2006 AQUACULTURE HEALTH INTERNATIONAL 19

new The first initiative, Aquaculture without Frontiers (AwF),
is an independent non-profit organization that promotes
and supports responsible and sustainable aquaculture and
alleviation of poverty by improving livelihoods in developing
countries. AwF does not seek to promote aquaculture in
isolation, but as a component of integrated rural and coastal
development plans, and of strategies to alleviate poverty (see
www.aquaculturewithoutfrontiers.org for more details).
The other three NGOs aim to support disadvantaged
communities in Africa or to take care of orphaned animals. A
South African-based non-profit NGO initiative, Animal Aid
for Africa (AAA), aims to uplift disadvantaged communities
across sub-Saharan Africa. Focus is on delivering sustainable
solutions to malnourishment and the spread of zoonotic
diseases, by exchanging skills and knowledge between
veterinarians. (www.animalaidafrica.co.za).
The other two organisations are Sophianum explores South
Africa, supported by the Dutch Ministry of foreign affairs
(www.ZA.DEBIOLOOG.nl and www.africanorphanage.
com), and Global Vets (www.ovc.uoguelph.ca/associations/
globalvets), a programme at the Ontario Veterinary College
that offers students a unique opportunity to investigate animal
health care in developing countries. Global Vets was formed
to promote international collaboration on animal health and
welfare, agricultural development and ecosystem health.
FISKERIFORSKNING INVESTIGATES
THE LINK BETWEEN SALINITY AND
SMOLT HEALTH
Irene Andreassen (Fiskeriforskning, Norway)
Normally, juvenile salmon grow up in fresh water, but in
today’s salmon smolt production, increasingly more seawater
is used. Scientists have now found that use of seawater before
the salmon smolt is ready for it can be detrimental.
Salt water is used in smolt production
for several reasons. Among other
things, it can help increase production
capacity and improve water quality in
some cases. However, incorrect use of
seawater can weaken the fish’s health and
might introduce pathogenic bacteria.
Our recent studies show that too high
doses and too early exposure of seawater DR HILDE TOFTEN,
during smolt production can result in SENIOR RESEARCH
reduced growth and additional problems SCIENTIST AT
with winter ulcers. Such outbreaks can FISKERIFORSKNING
impact fish welfare and have a negative
influence on the company’s financial return.
SEAWATER - NOT JUST “A PIECE OF CAKE”
It has been asserted that a gradual adaptation to saline water
before the salmon is fully smoltified is favourable, but the
research in this area has shown contradictory results.
In a Fiskeriforskning experiment, scientists chose to use
2 % salinity before the fish was fully smoltified, which is
not uncommon with smolt producers. “There is evidence
that such a high salinity isn’t particularly favourable for the
fish”, says Senior Scientist Hilde Toften at Fiskeriforskning.
Toften continued, “the fish ate and grew more poorly, and
became clearly more susceptible to winter ulcers. Our
results also show that these problems are aggravated when
we combine them with a very intensive production, which
has become relatively widespread in the industry in recent
years. So far, we haven’t known enough about this, but we’re
hoping that several factors will be clarified when the project
is completed.”
20 AQUACULTURE HEALTH INTERNATIONAL NOVEMBER 2006
The fish’s welfare is also very important for the industry,
and the Fiskeriforskning’s scientists are interested in studying
whether the practice of intermixing seawater affects the
salmon’s well-being.
“Of course, we can’t say anything with complete certainty
about how the fish experiences these problems, but we can
find the optimal conditions that ensure that the farmed fish
thrives, and which prevent the fish from becoming stressed
and sick. Winter ulcers are a serious disease which causes
large and deep wounds on the fish. If we can manage to
prevent outbreaks, we can help improve the welfare of the
smolt”, says Hilde Toften in closing.
The tests are part of a larger joint project amongst
Fiskeriforskning, NIVA, Norwegian University of Life Sciences
at Ås, University of Bergen, Marine Harvest and AquaNet.
The project is financed by the Research Council of Norway,
Marine Harvest, AquaNet and Fiskeriforskning.
For more information contact Senior Scientist Dr Hilde Toften
on phone +47 77 62 90 53.
INTERNATIONAL:
FROM BOXMEER TO DUBLIN!
Dr William Enright, the international marketing &
pharmaceutical manager for the Aquatic Animal Health
(AAH) Division, has decided to return to his native
country of Ireland. After nearly 11 years in Boxmeer, The
Netherlands, Dr Enright has accepted the position
of marketing manager for the companion animal & equine
business unit at Intervet Ireland. The move allows him to
return to Ireland and yet stay within Intervet. The change
in species focus and the more commercial environment
will offer him new and exciting challenges and learning
opportunities.
Dr Enright has been closely associated with the Intervet
AAH Newsletters. The first issue was published in October
2000 and it has appeared at six-month intervals since.
However, perhaps unknown to the aquaculture community,
his background includes research on a variety of animal
species and he has in excess of 250 publications to his
name. “It’s no wonder then that he has been an enormous
help to us here at Aquaculture Health International in
providing high quality articles and news items of interest
to our diverse readership,” says the editorial director, Dr
Scott Peddie.
“We thank William not only for his contributions to
Aquaculture Health International, but indeed to promoting
aquaculture in general. We will miss his expertise and valued
guidance, but wish him well for his continued success within
the Intervet family.” ■
WILLIAM
ENRIGHT
PROUDLY
DISPLAYING THE
FIRST TEN ISSUES
OF INTERVET’S
AAH NEWSLETTER
 BOOK REVIEW
SYSTEMIC PATHOLOGY OF FISH
A text and atlas of normal tissues in teleosts and their responses in
disease. Edited by HW Ferguson. Scotian Press 2006. 2nd edition
BY DR SCOTT PEDDIE. PATERSON PEDDIE CONSULTING LTD/AQUACULTURE HEALTH INTERNATIONAL UK

THE EDITOR
OF SYSTEMIC
PATHOLOGY OF
FISH, PROFESSOR
HUGH FERGUSON

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certainly not lacking in authorial gravitas. HIGH QUALITY PHOTOGRAPHS
At just under 400 pages in length, this ARE A MAJOR FEATURE OF THIS BOOK
publication is substantial. It is packed with
almost 700 extremely high quality figures 0ASTEURELLA
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ARCELLS • physiological and clinical pathology.
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Each of the above chapters contains a plethora

and electron micrographs. Indeed, it is no 16 of informative illustrations of both gross and
exaggeration to say that the quality of the microscopic pathology. The diagrams in the last chapter are also
photography throughout is second-to-none. A hardback binding worthy of particular note, given their “user friendly” and informative
and a relatively large page size further enhance the visual appeal. nature and their ability to dovetail with the text (which incidentally
The book consists of 14 chapters organised according to organ represents one of the best overviews on the subject I’ve read to date).
systems. The last chapter deals with physiological and clinical Most chapters start out with a description and a graphic illustration
pathology and is a new, and very welcome, addition to the second of normal organ morphology and function. Thereafter, specific
edition. A functional and aesthetically pleasing full-page colour abnormalities are discussed in depth and richly illustrated.
photograph separates each chapter. The chapter on the kidneys, written by Renate Reimschuessel and
The introduction sets the scene nicely for the rest of the book Hugh Ferguson, is a case in point. It opens with an illustrated section
by summarising post-mortem techniques and general pathology. on general anatomy, and then looks in some depth at the response of
The writing style is very fluid and makes for an easy read, without the nephron and interstitium to disease, and closes with a discussion
compromising any of the detail required in a textbook format. on the effect of parasitic infection on kidney function and pathology.
Subsequent chapters deal with Hugh Ferguson’s book will appeal to a wide range of researchers,
• gills and pseudobranchs diagnosticians and students. It is readily accessible to those familiar
• skin with fish biology, but not necessarily experienced in pathology. That
• kidney said, it will also be an invaluable reference tool for pathologists and
• spleen, thymus, reticuloendothelial system, blood diagnosticians more used to working with other species.
• cardiovascular system All in all, this book is an excellent, highly informative and well-
• gastrointestinal tract, swim bladder, pancreas and peritoneum balanced addition to the somewhat limited range of fish pathology
• liver textbooks currently available. It is one of those rare publications
• nervous system that can only be described as a “must have” for any serious fish
• the eye health professional.
• endocrine and reproductive system For sample pages, and ordering information,
• neoplasia, and as mentioned previously, see www.scotianpress.com ■

NOVEMBER 2006 AQUACULTURE HEALTH INTERNATIONAL 21

FAC I L I T Y F O C U S
FISH HEALTH INVESTIGATIONS AT
ARDTOE MARINE LABORATORY,
SCOTLAND BY DR JIM TREASURER
ARDTOE MARINE LABORATORY
LARVAL
REARING TANKS
A
lthough ownership of Ardtoe has changed recently and the
company trades as Ardtoe Marine Laboratory, research on
fish health continues. The site was well known as the Marine
Farming Unit, with a remit to develop methods for cultivating
marine finfish following its inception in 1965 by the White Fish
Authority, later the Seafish Industry Authority.
Work was initially on plaice in an extensive lagoon system, but
later on turbot. From 1983 Ardtoe pioneered the culture of halibut
in the United Kingdom, and this was followed by wrasse culture
and then demonstration projects with Marks and Spencer and other
partners on cod from 1998 and haddock from 2001.
The site currently has a wide range of fish broodstocks, including
cod, haddock, halibut, dover sole, turbot, whiting, pollock, saithe
and wrasse.
FISH HEALTH
Survival of marine larvae in culture has been low, for various
reasons, and much of the research at Ardtoe has focussed on fish
health and welfare issues.
Vibrio anguillarum is recognised as the main threat to cod and
haddock farming. Work at Ardtoe, in association with KTP fellow
Yolanda Corripio of Aberdeen University, has looked at the immune
22 AQUACULTURE HEALTH INTERNATIONAL NOVEMBER 2006
KTP ASSOCIATE YOLANDA
CORIPIO STUDIES THE
DEVELOPMENT OF THE
HADDOCK IMMUNE SYSTEM
system of gadoids and the development of suitable vaccines.
The optimal route to challenge haddock was investigated with
maximum effect by immersion. Five vaccination regimes were used
and efficacy tested by bacterial challenge. There was a significant
decrease in mortalities compared with unvaccinated controls. As
in previous cod studies, vaccination of haddock did not induce a
specific antibody response.
DISINFECTANT STUDIES
Larval rearing systems have a low exchange rate to reduce the loss
of live feed from the system and this, together with the use of live
feed such as rotifers and Artemia, can introduce a range of bacterial
species, including Vibrios.
Disinfection of eggs and reducing the risk of transferring
bacteria to the rearing tanks has been examined using short-
contact immersion in Pyceze (50 percent w/v bronopol) (Figure 4),
commonly used in salmonid rearing.
Static immersion in the rearing incubators for 30 minutes and
exposure to 50mg l -1 bronopol also gave improved survival compared
with controls and significantly reduced bacterial numbers.
Pyceze was also compared with ultraviolet in the disinfection of
live feed by addition during the enrichment process of live feed. A
 FIGURE 4: SURVIVAL OF COD EGGS FOLLOWING CONTACT DISINFECTION FIGURE 5: SURVIVAL OF COD LARVAE TO HATCH IN THREE TANKS SUPPLIED
WITH BRONOPOL FOR 45 SECONDS WITH LIVE FEED TREATED WITH PYCEZE, COMPARED WITH THREE REARING
UNITS WITH UNDISINFECTED FEED
Survival of cod eggs to hatch

survival at 75 days
Percentage larval
100
15
% survival to hatch

80
12
60
9
40
6
20
3
0
50 Bronopol 500 Bronopol 1000 1000 Seawater 0
Bronopol Kickstart control Control Disinfected
Treatment/conc mg/l
Live feed treatment

suitable concentration and contact time to disinfect live feed during A COD LARVA WITH INGESTED SIBLING
the enrichment process was established as 30mg l -1 active Pyceze for (PHOTO COURTESY OF HEATHER FORBES)
six and 12 hours. This was tested on a commercial scale in six 1300
litre larval rearing tanks with higher survival in larvae fed disinfected
feed, with an average of 13 percent from egg to weaning in three
experimental tanks, compared with seven percent in control tanks.
(Figure 5)
Following this work, Pyceze is now routinely used for disinfecting
live feed in many marine hatcheries worldwide, and work continues
in looking at other applications of Pyceze in marine hatcheries.

COD AND HADDOCK WELFARE

As cod and haddock are naturally cannibals, large losses and fin
damage can occur in hatcheries if grading practices are not rigorous. importance of ectoparasites. Recent
A research student from Glasgow University, Heather Forbes, is support from Seafish and a salmon
working at Ardtoe, and has focussed on how the size of prey items company has primed a rearing
can affect the development of the jaw apparatus, with large prey programme for certified disease-
items inducing larger gape size, encouraging the development free wrasse to stock with farmed
of cannibalism. Results will be used to recommend feeding and salmon to control sea lice.
management practices that reduce cannibalism. While cleaner fish have been
As various health issues of gadoids are encountered, investigation used previously in salmon
continues on subjects such as the control of atypical furunculosis to farming, there have been HUSBANDRY
IMPROVEMEN
eye problems, the latter especially in halibut. Aggression can cause concerns in using wild-sourced ARE TACKLIN TS
G EYE DAMAG
E AND
damage or eye loss, and cataracts may occur associated with diet and fish, and the trend to organic CATARACTS
IN HALIBUT
UV exposure, and these are also being investigated at Ardtoe. production has awakened interest
in the use of wrasse in sea lice control.
DIET RESEARCH While Ardtoe increases the production of cod juveniles for
Nucleotides are considered as semi-essential nutrients during food diversification of marine finfish culture and to cover the core costs
deficiency, rapid growth and immunological stress. In a study of operating the site, further work will be on key health issues
by Jose Vecino in association with SAMS, broodstock diets were affecting the health and welfare of marine finfish species. Dedicated
enhanced with nucleotides in halibut and haddock. This was found trial facilities and an on-site residential accommodation unit have
to improve fecundity, egg quality and fertilisation rates in both just been completed to encourage students and visiting researchers
species. The effect of diet enriched with nucleotides was also studied to enjoy the benefits of working in the relaxing environment of the
in cod larvae with Yolanda Corripio. spectacular Ardnamurchan Peninsula.
As the cost of marine ingredients and the availability of fish oil and For further information, visit www.ardtoemarine.co.uk ■
meal will be potentially restrictive to the expansion of aquaculture,
Ardtoe has been carrying on trials with IFFO and various feed
companies on the effect of partial full-fat soya substitution on cod
performance, and also looking at the anti-nutritional effects on cod
growth and potential effects on the gut lining.
While the liver has not been affected, there have been minor
effects such as a moderate inflammatory cell presence in the
posterior gut, although these have been found to be mild compared
with the effects of vegetable diets in salmonids. It also appears that
after an initial induction period of several weeks, cod and haddock
adapt to the addition of vegetable protein in the diet.

ONGOING WORK MILD INFLAMMATORY RESPONSE IN THE POSTERIOR

GUT OF COD FED DIET WITH FISH MEAL AND OIL
Project work with ongrowing partners continues on the growth
PARTIALLY SUBSTITUTED WITH VEGETABLE PROTEIN
and health of cod and haddock in sea cages, including the potential

NOVEMBER 2006 AQUACULTURE HEALTH INTERNATIONAL 23

DIAGNOSTICS

DIAGNOSTIC
LABORATORY SERIES
SERIES EDITORS: DR DAVE GROMAN AND DR FRANCK BERTHE
(ATLANTIC VETERINARY COLLEGE AT THE UNIVERSITY OF PRINCE EDWARD ISLAND, CANADA)

A
variety of service, research and teaching laboratories exist
worldwide which support the aquaculture industry. These
laboratories often offer disease screening and diagnostic
services, with various levels of testing and quality assurance.
In addition, some laboratories may not engage in pro-active
international marketing. As a result, many aquaculture companies
and their fish health service providers are not always aware of the
range of laboratory resources available in the global marketplace.
This series of laboratory articles will provide Aquaculture Health
International readers with a guide to diagnostic laboratories which
offer regional, national or global “routine - fee for service” veterinary
diagnostic services to finfish, mollusc and crustacean producers and
their veterinary service providers.
In addition, the articles will focus on affiliations that these
diagnostic laboratories may have with universities, government
agencies and institutes linked to aquatic health training or research.
The articles will seek input from each laboratory as to their strategic
goals and operational philosophy.
This objective services review, in combination with subjective
input on management philosophy, will provide readers with a
balanced description of the laboratory, and will ultimately help
aquaculture veterinary professionals to make informed decisions on
selecting appropriate diagnostic service laboratories, aquatic health
training and research programmes.
To accomplish this, we will provide a formative review of
the services provided by each laboratory, with the approval and
assistance of the company, programme or laboratory management.
To this end, we have developed an aquatic health diagnostic services
evaluation checklist which will detail information on the type and
scope of services offered:
• quality assurance programmes
• referral options
• reporting methods
• client base, and
• the cost of testing.
We will strive to capture a thorough description of the diagnostic
component of the laboratory, with a capsulated summary of services
provided. If a laboratory prefers not to participate in the series, we
will only provide a description based on published information,
public advertising or government documentation.

FEATURED DIAGNOSTIC LABORATORY:

Veterinary Diagnostic Service (VDS),
Institute of Aquaculture, Stirling University, Scotland
BY PROFESSOR HUGH FERGUSON
infections in farmed cod and

T
he Veterinary Diagnostic Service (VDS) is located within the sole, and reoviral-like hepatitis
Institute of Aquaculture at Stirling University, Scotland. It was in farmed halibut.
established in 1972 and is committed to providing a dedicated The Veterinary Diagnostic
and leading role in the support of global aquaculture, fisheries and Service has access to laboratory
aquatic animal health in general. capabilities covering a wide
We work with individuals, companies and organisations to range of disciplines. The service
develop and provide total health management, ensuring high also has access to all the other
standards of veterinary care and attention. expertise, information and
Staff at the VDS Stirling work mainly in a consultancy and referral facilities within the Institute SAMPLING A
SALMON IN
basis on samples sent by veterinarians, fish farmers and companies of Aquaculture. Currently the THE PATHOLO
GY LAB
(feed and pharmaceutical industries), and through other research VDS has two clinical veterinary officers,
organisations, to develop and provide advice and guidance on with Dr Ferguson as director and chief pathologist. The service
health management of research, farmed, wild and companion is supported, either directly or through arranged consultation, by
aquatic animals. the Institute of Aquaculture staff, which includes five additional
Definitions of new syndromes and pathogen identification professors, 14 associate professionals, 30 technicians and 25 research
continue to be the strengths of the VDS Stirling. Many new diseases fellows.
have been identified over the years, and these have formed the
basis for a large number of MSc and PhD research projects within SERVICES PROVIDED
the institute. Professor Hugh Ferguson, who joined the faculty in VDS at Stirling operates on a cost-recovery basis, and has supplied
1998 to head the diagnostic services, introduced a rapid 24-hour a diagnostic and referral service for decades, evolving over time to
turnaround in disease diagnosis, much faster than most mammalian provide rapid diagnosis of aquatic animal diseases caused by a wide
diagnostic facilities! variety of pathogens affecting a diverse range of farmed aquatic
This rapid histopathological service includes provision of animals. The full spectrum of diagnostic, research and consulting
management strategies to improve the health status of a wide range services is available, including:
of fish species, and is fully supported by active and experienced • Histopathology
technical staff. Examples of some recent local diagnostic “firsts” • Bacteriology
include VHS in farmed trout in the United Kingdom, nodaviral • Virology

24 AQUACULTURE HEALTH INTERNATIONAL NOVEMBER 2006

THE ELECTRON
MICROSCOPE IN
OPERATION

ONE OF STIRLING’S
WELL-EQUIPPED
LABORATORIES

• Advanced molecular techniques pharmaceutical companies.

• Electron microscopy • Veterinary surgeons, where referral work and laboratory service
• Parasitology for veterinary surgeons in practice is provided and is also available
• Serology to aquarists, fish wholesalers and public aquaria.
• Immuno-histochemistry
• Health certification COLLABORATIONS AND LINKS
• Veterinary investigations Staff in the VDS Stirling not only provide a diagnostic service, but
• Health screening also are also involved in research into new and emerging disease
• Specific pathogen testing problems in farmed fish species. The VDS also participates, through
• Site visits the institute’s Aquaculture Technology Centre, in commercial
• Feed analysis
• Water chemistry
• Pathology referral service
• Expert reports
• Training
• Contract research, and
• Market research
For example, bacteriology workups include isolation and
identification of fish pathogens, morphological, biochemical and
molecular identification techniques and assess to the VDS reference
bacterial fish pathogen collection.
Similarly, virology testing is done by a dedicated virology unit
with cell culture and virus isolation facilities with access to a wide
range of cell lines and molecular diagnostic techniques.
Some services are provided to a quality assurance level of good
laboratory practices, a fact that vaccine and pharmaceutical
companies find very useful for getting their products to market.
Although the laboratory is based in the UK, the service provided
is global. For example, the first description of a bacterial disease
causing mass mortalities in farmed catfish in Vietnam, Pangasius
hypophthalmus, was reported and the aetiological agent identified
as Edwardsiella ictaluri. VDS include the following categories of
clients:
• Aquaculture companies, where the VDS provides veterinary
assistance and advice is provided to aquaculture companies
through farm visits, regular fish stock screening via parasitology,
bacteriology, histology, virology and specific pathogen certification.
Integrated health management contracts are presently established
in Scotland, England, Ireland and Greece.
• Nutritional, pharmaceutical and insurance companies, where
veterinary investigation for nutrition and insurance companies
is regularly provided, through either retained veterinary
services or specific investigations. Expert reports, market
research, field trials and contract investigation are provided for

NOVEMBER 2006 AQUACULTURE HEALTH INTERNATIONAL 25

RESEARCH
SITE VISITS ARE AN
IMPORTANT PART OF
THE SERVICE OFFERED
AT STIRLING
THE STIRLING
enterprises on a worldwide basis, by
offering a range of contract research, analytical and knowledge-
based services that will help companies to innovate, improve
efficiency and competitiveness, enhance product quality and ensure
sustainability.
The merging of disease diagnosis and research is very
strong within the VDS, and has enabled more sophisticated
diagnostic techniques to be performed. Application of immuno-
diagnostic and molecular tools (including sequencing)
has enhanced identification and confirmation of viral and
26 AQUACULTURE HEALTH INTERNATIONAL NOVEMBER 2006
bacterial pathogens of significance within
the aquaculture industry.
Combining molecular tools with routine
histopathology is a particular aim of the
VDS and, together with advanced pathogen
identification techniques, these tools have
conspired to markedly improve the provision
of practical management strategies to alleviate
aquatic health problems globally, thereby
enabling the VDS Stirling to continue to
provide high standards of veterinary care
and attention.
The research focuses on identifying the
aetiological agent(s), understanding the
disease processes and associated influences
on disease outbreaks within fish farming
systems in the UK and Europe. Information
IVERSITY CAMPUS from all research is widely disseminated to
UN
the fish farming industry through various
media. Research tools are then applied
within the diagnostic facility to strengthen pathogen identification
and disease diagnosis capabilities to help promote sustainable
aquaculture practices.
Research is currently being conducted into cardiomyopathy
syndrome of farmed Atlantic salmon, as well as into two new
syndromes affecting the rainbow trout industry: red mark syndrome/
cold-water strawberry disease, and rainbow trout gastro-enteritis.
Contact Professor Hugh W Ferguson, email h.w.ferguson@stir.
ac.uk, Richard Collins (r.o.collins@stir.ac.uk) and Dr Mags
Crumlish (mc3@stir.ac.uk), or see www.aquaculture.stir.ac.uk/
diagnostic/index.html ■
NOVEMBER 2006 AQUACULTURE HEALTH INTERNATIONAL 27
RESEARCH

PROBIOTIC BACTERIA’S
EFFECT ON SHRIMP SURVIVAL
BY MATHIEU CASTEX (IFREMER, NEW CALEDONIA), VINCENT USACHE AND SYLVIE ROQUEFEUIL-DEDIEU (LALLEMAND, FRANCE)
ALL PICURES COURTESY OF IFREMER

S
hrimp production in New Caledonia is rapidly growing, important amounts of lactic acid
with production aiming at 5000 tonnes per year by 2010. (homofermentative strain). It is
Production is essentially destined for export, mainly to France, also thought that P acidilactici MA
Japan, Australia and the United States, and for this reason the sector 18/5 M prevents pathogens from
must keep up with the demand for quality and high added value attaching to the gut surface and
products, in line with international safety standards. colonising it, by competition.
To attain these objectives, pharmaceutical solutions must be kept Previous production trials have
to a minimum and natural, scientifically proven preventive solutions shown the zootechnical effects of
must be sought. In this context, the use of probiotics seems a Bactocell on marine shrimp. For
promising strategy, acceptable by consumers, and scientifically backed example, a trial conducted at the
up to promote shrimp growth and protect them against infections. Vung Tau Centre for Shrimp
This article presents the first results of a joint research project Research and Production,
between probiotic specialist Lallemand and Ifremer (French in Viet Nam (2002) showed PROBIOTIC STRAIN
PEDIOCOCCUS
Research Institute for the Exploitation of the Sea) which was that continuous Bactocell ACIDILACTICI MA
18/5 M
initially presented at the WAS Aqua 2006 Conference. supplementation in feed of juvenile
This project shows the benefits of probiotic lactic acid bacteria Penaeus monodon:
P acidilactici MA 18/5M administration on shrimp zootechnical • improved growth rate by 52 percent after eight weeks
performance and survival following opportunistic infections, • improved size homogeneity, and
particularly “winter syndrome”. • increased shrimp length by 15 percent.
This effect on growth could be linked to improved feed
PROBIOTIC CONCEPT digestibility.
Microorganisms are commonly used in aquaculture to control
microbiological ecosystems, especially to treat the water in tanks WINTER SYNDROME
and ponds. This is known as the bio-remediation concept. The winter syndrome, or syndrome 93, has had an important
More recently, the addition of live microorganisms through impact in New Caledonia, as it has been responsible for very high
the feed (the probiotic approach) to balance the animals’ gut mortality rates (about 60 percent). As a result, most shrimp farmers
ecosystems and improve digestion and sanitary conditions has been have had to stop production during winter, resulting in significant
attracting growing interest. The advantage of probiotic over bio- economic losses.
remediation is a direct effect of the micro-organism in the gut, due The winter syndrome is caused by Vibrio penaeicida. When the
to its ingestion, as well as a better control of the number of micro- water temperature decreases below 22° C, shrimp are subject to
organism delivered to the animal and the environment. physiological perturbations and then become more vulnerable to
In particular, the probiotic strain Pediococcus acidilactici MA 18/5 the pathogen present in the water.
M (Bactocell®, Lallemand, France), which is a lactic acid bacteria, In the trial conducted at Ifremer in Nouméa, Litopenaeus stylirostris
has been specially selected for its ability to strengthen the gut shrimp reared in tanks received Bactocell in the feed twice daily (two
microflora of monogastric species, with a positive impact on both doses were tested: 106 and 107 CFU/g feed*) during the cold season.
pathogen control and feed digestibility. Mortality was recorded daily (Figure 1). The probiotic treatment
It is characterised by its ability to produce exclusively and rapidly significantly improved the shrimps’ survival.

FIGURE 1: DAILY MORTALITY AND TEMPERATURE VARIATIONS IN TANKS FIGURE 2: OVERALL SURVIVAL AFTER 31 DAYS REARING IN TANKS

Survival rates among treatments after rearing

Survival %

27
Temperature °C

100
31 days in tanks (n=9 for each treatment)
26 120

90 Bactocell 10^6 CFU/g

25
100 Bactocell 10^7 CFU/g

24 +42%
80
Survival (%)

80

23 +23%
60

22
70
40
Control
21 Bactocell 10^6 CFU/g
Bactocell 10^7 CFU/g
20
20 60
0 5 10 15 20 25 30
Rearing days 0

28 AQUACULTURE HEALTH INTERNATIONAL NOVEMBER 2006

 THE EXPERIMENTAL TANKS WERE
SUBMITTED TO TEMPERATURE
VARIATIONS TO INDUCE WINTER
SYNDROME IN SHRIMPS

When decreased water temperature lead to nearly 50 percent
mortality in control tanks, the survival for probiotic treated shrimps
was on average 75 and 95 percent, with 106 and 107 CFU Bactocell/g
feed, respectively. There was a significant dose effect. Globally, after
31 days treatment in tanks, the shrimp survival rate was increased by
up to 42 percent with the higher dose of Bactocell in the feed.
Several mechanisms have been put forward to explain these
beneficial effects of probiotic, although on-going studies are aimed
at further elucidating the mechanisms involved in the case of
Bactocell in shrimps. These include:
• the competition or exclusive competition for space at the gut
CONTINUED ON PAGE 31

GUT MICROBIOLOGICAL STATUS

Further microbiological analysis of the shrimps’ gut was performed
in order to estimate the effect of the probiotic on the bacterial
gut microflora. The total number of bacteria and the total vibrio
population were assessed in the gut of shrimps from the control
group, and the group receiving the higher dose of Bactocell (106 and
107 CFU/g feed), at day 16 and 30.
It appeared that both the gut total bacteria and the vibrios
populations were significantly inhibited with the probiotic
treatment. These results indicated that Bactocell could improve
shrimps’ survival against pathogenic infection by acting on the
gut microflora, in particular by reducing the number of vibrio
pathogens in the gut.

FIGURE 3: SHRIMP GROWTH RATE IN TANKS IS IMPROVED WITH DAILY

ADMINISTRATION OF BACTOCELL

Mean daily growth rate of shrimp reared 31 days in tanks

0,065

0,060
Growth rate (g/day)

0,055

0,050

0,045

0,040
l /g /g
ntro FU FU
Co 6C 7C
el l 10 el l 10
c c
c to cto
Ba Ba

NOVEMBER 2006 AQUACULTURE HEALTH INTERNATIONAL 29

PERSPECTIVES

COARSE FISHERIES AND

FISH HEALTH IN ENGLAND
BY BERNICE BREWSTER (AQUATIC CONSULTANCY SERVICES, UK).

M
ore people in the United Kingdom
enjoy the pastime of angling than
any other sporting activity. The
tradition dates back many centuries as
evidenced by Izaac Walton’s treatise on
British fishes in 1653. Freshwater angling is
administered by the Environment Agency,
a government agency responsible for
the licensing of the sport, and sales of
rod licences exceed 1.2 million annually.
Licences are also sold for game fish, but
these are not included in this figure.
Coarse fishing currently attracts the
greater interest.
In the middle of the last century, most
coarse fishing was largely confined to
rivers or canals, with a limited interest
in the lakes or still waters. Carp (Cyprinus
carpio) were and still are revered for their wily nature and difficulty A CARP SUFFERING FROM ERYTHRODERMATITIS
to catch. The 1950s saw an upsurge in fishing still waters for carp, In the early 1950s, when Clarissa was caught, such a large carp
with the legendary angler Richard Walker setting the record with a would indeed have been unusual for this country because the
specimen later named Clarissa, that weighed almost 20kg (44lb) in weather conditions were not ideally suited for rapid growth.
1952. Richard Walker’s capture of Clarissa was the spark which ignited
Subsequent to the publicity surrounding Clarissa’s capture, the the passion for carp in the UK, with this species increasingly
passion for carp fishing in the UK has increased year on year. These attracting more attention. In the latter part of the last century, a
days it is probably the still waters that attract more attention to “double figure” (ie a specimen weighing more than 4.5kg or 10lb)
anglers than the abundance of beautiful rivers in this country. An carp and possibly a “twenty” (9kg) was regarded as a good catch.
increasing number of lakes are being excavated, and as agriculture Regrettably the trend to want to catch ever-larger carp continued,
and farming continues to suffer from poor returns, more farmers which resulted in the illegal importation of large, specimen carp
are diversifying by creating angling lakes.
It is worth bearing in mind that although there is some discrepancy
as to the actual date when carp arrived in this country, they are not Many carp are known as
native. After the initial importation of carp they were indeed on the
extreme of their range,
“grinners”, where repeated
being a native to the catching has damaged their
Mediterranean region,
naturally occurring jaws and tissues, resulting in
in slow-flowing rivers
or lakes where the seriously deformed mouths
water temperature is a
minimum of 18˚ C. from continental Europe, sometimes accompanied by outbreaks
If indeed the of the notifiable disease spring viraemia of carp virus, or SVC, into
favoured date for which these imports were stocked.
introduction is the The laudable work of the Centre for Fisheries and Aquaculture
mid-16th century, the Science, which managed to catch and prosecute many illegal carp
F “GRINNERS” climatic conditions importers, was later supported by the formation of the English Carp
EXAMPLES O
were unfavourable Heritage Organisation, which has achieved an excellent following,
for this species and the trade of illegal fish is slowing. Certainly, UK farms produce
and perhaps it is large numbers of carp for the angling market annually.
surprising that However, the popularity of carp is not waning and seems to
it managed to go from strength to strength. As a result, this species has been
survive. None-the- stocked into almost every still water in the country. Certainly
less, survive it did, some 48 percent of my annual business is examining carp for
although growth movement consent.
and reproduction Coupled with the desire to catch specimen fish is the onset
would have been of a society which demands instant gratification, and angling
severely limited. is no exception. As a result, many fisheries are heavily stocked,

30 AQUACULTURE HEALTH INTERNATIONAL NOVEMBER 2006

predominately with carp, but also bream (Abramis brama), roach
(Rutilus rutilus), rudd (Rutilus erythrophthalmus) and smaller
numbers of other species.
The numbers of fish stocked into fishing lakes that may be either
commercial ventures or even those administered by angling clubs
and societies can exceed 2000kg fish weight per hectare. Such
intensive stocking of fish impacts on the aquatic environment, with
little in the way of submergent or emergent plant life.
Fish stocks quickly destroy the submergent plants, which can
never thrive because of the discolouration of the water. The knock-
on effect is that there is little in the way of natural food for these fish,
which become entirely reliant on anglers’ bait for nutrition.
There are two issues with regard to this matter. Firstly, most
angling takes place in the warmer summer months, but once
temperatures cool, the numbers of fishermen and therefore
sufficient food for the autumn and early winter when the fish are
still feeding diminishes. Secondly, while anglers are keen to use
baits with attractants and stimulants that draw the fish to the bait,
there must be a question over the nutritional completeness of such
a diet and its shelf life.
HEALTH IMPLICATIONS
Given that carp are the preferred species in these heavily stocked
waters, the fish are subject to tremendous angling pressure. Many
carp are known as “grinners”, where repeated catching has damaged
PROBIOTIC BACTERIA’S EFFECT ON SHRIMP SURVIVAL
CONTINUED FROM PAGE 29
surface, the production of molecules inhibiting
the growth of pathogens, such as bacteriocins or
antibiotics, or
• creating a hostile environment, such as low pH (eg
lactic acid production).
Finally, the new trial also showed a significant
improvement of shrimp growth performances, with
a dose effect. In tanks, growth rate at 31 days was
improved by 10 percent and 16.5 percent respectively
(Figure 3). These results confirmed previous studies
done in commercial rearing conditions,
and demonstrated the economic
relevance of using probiotics in shrimp
production.
PROBIOTIC PATTERN
As seen before, the modes of action of
probiotics such as Bactocell is linked
to their activity, implying that in order
to exert their effects, probiotics must
remain alive until they reach the gut. For aquaculture applications,
in particular, this means that the microorganisms must survive
the stressful conditions of pellet feed processing (particularly
harsh conditions in case of shrimp feed, with exposure to high
temperature and pressure), feed storage, the aquatic environment
and the digestive process.
For these reasons, extensive studies were conducted to determine
the optimal way of incorporating Bactocell into shrimp feed to
ensure its survival up to the animal gut and to guarantee the
expected count of live cells. The method finally selected was a post-
pelleting incorporation, in fish oil.
Bactocell showed good stability during the process, and the feed
could be stored for up to 15 days at 25˚ C. The probiotic survival
was still satisfying after 48 hours immersion in marine water. Finally,
their jaws and tissues, resulting in seriously deformed mouths, and
often you have to wonder how on earth they manage to feed.
Clearly such damaged fish show poor growth due to the restraints
on feeding, which perhaps makes them ever more likely to be caught
as the angler’s bait becomes an easier meal. Over a period of years
those that are the most severely deformed probably simply lose
condition and die.
It is also common to find carp suffering from carp erythrodermatitis
on heavily stocked or fished waters. These ulcers are often found on
almost every fish that is caught.
Treatment is impossible because the underlying stressors of poor
nutrition and repeat catching cannot be removed. These of course
are just the physical manifestations of a heavily stocked fishery, and
the stocks suffer from the same physiological effects of an intensive
culture system.
The difference of course is that in intensive culture the fish
are culled at a certain size, usually for the table. It is also worth
remembering that intensive stocking of fish and its associated stress
provides the best background for the emergence of disease.
Carp are traditionally regarded as being strong fish, possibly
because they appear to tolerate handling, and they are certainly able
to survive in polluted water and low oxygen. But they easily succumb
to a range of diseases, some of which, such as spring mortality of
carp syndrome, are of unknown aetiology. Perhaps they do not
tolerate handling and poor conditions as readily as we assume. ■
INDIVIDUAL SHRIMP MARKING
ALLOWS INDIVIDUAL WEIGHT
MONITORING FOR MAXIMUM
STATISTICAL RELEVANCE
Bactocell resisted well the digestive process as shown by
live bacterial count in the shrimps’ digestive tract. The
experiment also confirmed that Bactocell did not colonise
the animals’ gut. It is for this reason that it has been administered
daily, with each meal, to ensure consistent effects.
CONCLUSION
Lactic acid bacteria is a natural solution to improve shrimp
performance and to ensure producers receive a good return on
investment. Most importantly, it represents a good approach to
controlling gut microflora, keeping pathogens at bay and improving
survival in conditions of stress.
*CFU: Colony forming units. The measuring unit used to count live
microorganisms. Only microorganisms that are able to multiply on a
Petri dish (and form a colony) are taken into account. ■
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32 AQUACULTURE HEALTH INTERNATIONAL NOVEMBER 2006